·THE NEXY TURK ACADEBAY OF SCIENCES (Forcad3317) COUNCIL, 1959 Froldant ILURY KOPPONE President 2 L) XOPAC Vkehalders LIZE PORT CP RIQADS frordikt Secretary Corra pondoh[ Sennert 2. EILSON DAY FREDERICK 125ELOBLE Treanor GUSTAV I KARTIN Elded Councury I 1999 POTGI I LYOUN HAYDD, MICHOLSON SITEES VULLIALI W VALLOTT 1951-1960 041 VIFAR.OFSKY CUTTAV I LAPTT HTL#CALAMIT FORIN E VAN NU I DETTRICK QLARLI W BOSJETI 1 BELISOKIS EL TATUMS Fimona Content TLARDES I TAYLOR, Chinos CORDOBILLARD ROBERT T LIGUT Ermiry Drede FL TCE THOMAS SISTER sicTJOY OP DJALOOCAL ARD XEDICAL STVU TOIRT LLOC, GAN CHARLES NOBACX Taruhan DUTINIO OF LATA POMOLOOT IX ROTTY I CUP ry DOROTA CROSS JEXSIY Serwony Di Varax or 146TRUXIST TJOY DI \ NO C ANDRES TERRARL Sporty i otro. I curoor SARL UIRANIOPOSETL, 7 mm EVATTIL GRUDIRC, Sur bir OY or rejOLOGY GreCM RVRAS Chow VICTION OF CAPRICAL ACII CES TAKL FOLKERS Car FREDERICK R INRICTL, Story FRETTY OF Otological ARU SURT IMIT CA YREDERICK N YOUNG Jr Story pymap of OJNO PAT AD XTTO door ALRALA I SUSSL Cher CJLARLES CAUDIN Stonwery ArctOY A POCAL CYCTJ E.147 LTLR FIORI, Clair BROVHERO Story QTISOY OF SCOUT TU OSD DSL4 Cut Product TUU E GRFTON NOBLACE W STLYIRD LARDO P TAYLOR VICTOR LAXER NL CROSS DORA REGEL OSS 1GXLUI The Divky Hraunty samoch det iz Ort Vlsy, tacting Aneh dah bakal The Nr Away #ifa synd Fut Dr IL, Ks Cacte em bist mit kleriku tun tatoved by you ·ANNALS OF THE NET YORK ACADEJTY OF SCIENCES HOLOWE 2 ART 12 PAous 387 558 February 6 1959 Ed lor in Clad OTTO Y SE HITLOG laxaging Editor FRAXILDI N FUNT amale Eduar JardID. SELEX METABOLIC FACTORS IN CARDLAC CONTRACTILITY Cows xp Editor ALENARD V GELA CONTENTS 390 Introductory Ram 5, VDA N Goran 09 10. ipd Ther Relations to Hart Monche Genel Suvcy d Ion Cocotton by Cronor C Gum laintrance of Concentrato ndikots and Regulation of Cd Volume By ALDTADA LEAL 326 Phywopal Effect of lope Dy RODROM 09 Elint of Toon on the Actmty of Ency per Derned frog Cart Two D JONA Mur 415 Intention of Chycolyes and Dadatie Puth BE RAIO AD 8 CATT Da Gyong Storer th Speal Redentot tbe Cardoc Type of Gentuted Glycoporowy PAUL A D Sac horou The Role of Co, Foton o Carbobbat Utitombok od Sydens By Maroor UTID 451 Eneo-Goneratny dydet By SATILLO GIRLIA +02 Trematon of Emp I Canbau Wot By Roro E OLOT DOTAYA PUTIO 466 Electro Trunport Card Two By Tonus P SINCE Omdative Pbosphoryhtyn o Hudo Mitochondon and Iu Rubik Rubce Putr Survitz 30 Oulator Prophoryleboo Storbo ho Voral and Eppuretat Protoond Cognitive Heart Tulon m Guna Php Compano BiGWE PULT NUD 1 Gen 513 Hyachrooy d Suncten und Foctual Canon Carte la litochondo Lan P. 518 The Choral Tbomoh mo a Votorider Hedino o Vaclar Cootruction By RJ Popuu lycke Proton Croy Ubbatio BJ Goon Aartha Fomcy JR Jong O UDAL 15 Too oppe the rear of aeon Vettel last CAR Cat Altyd by The No Yort ladeoy Boxca Verd 1: od 19 1 ·ܝܢ ·INTRODUCTORY REMARKS Verard J1 Catler furthed Uddin ow Doteka, Perry-BAW Voel Cruto Fairy In recent yans attention bar beca focused on the etiologtaal lectont contributing to chronic bean doak, that the bypertense, atherosclerou thematic, and congenital lacton The terminal term problema sa the absence of sudden death) often congestive heart fullore For various retom it la dificult to know at the present time the exact madeace of congestir bart abort A bogpuat ward Airvey and a parvey of 1000 consecutive private por trent de doud (1) 31 ans of grada 3 to congestire fatture in 180 ward bedie od (2) 93 private an oth grade 1 to 2 coopertine falure (no attempt us made to cut the etiologial type of beart dorense) The terminal espect of beurt del crantes high degree of morbodity in our popuation. It a difficult to determine at present whether pathological trutural changes 10 the myocardiom precede bundemical change, ibat , phycochemical alteration that may result in onk imbalance, mkrastructural aberration, or proteta low that then produces a pattern of Xequental estate terminating in arduc luhure. Its nuumed, us a working hypotbess, that we of the principal and fidel od of congestivo bart failure un creence a fult of the arulable chem Koal cery to be inankorroed and otulued u mecranual energy by the bear The purpose of the modo upo s to generate and promoto new conceptu concerning concertire beurt failure that may be amenable to ermination by techarques hitherto unxed in this perimeter In addition, it is thought that of the problem of congestive heart fulure were presented by those familias eith the problem to odrudub of other dociplina who tre upucquanied with the problem, net approaches for Hudy mybt become spparent Accordingly the conference on wbxcb thus monograph w baxed nu derned to bring lorather comun phrralagets buocbemuts, bophyunt, add phryants in order that they might dorm the problem of congestive bear falare from the diretto view pounts. The con lertilization of post so y proach has proved to be of madmato value in other wodies, and ac hope that it Ol prove to be vtruable in the bed a mer ·JOS A THIEIR RELIGIOSHIN TO HEART MUSCLE I CITRIL SCRIFI Of 10 CONCENTRATIO (corge (otith De VNIK (ITH Sad CH Ark SchedVakiw LA tamo Chidy Und Led to Call In 1954 I joined Butt eod llwer of the Univerity of Southern California School of Medicine in the publeton of data on the oncentrations of copper uron, lad mentury manganese and zinc in various organ of the body Our data had been obtained through glettrographic and chemkal mlyns of troues talen al autony from 910 patients stored at the Los Angeles County Hor mual Investigation into the physiologial and pathological wynitrance of inue metals in man was getting under way at that time and there wu need for mirmation on the normal levels of each metal in venous body times, In order that we might go on to klentify and investigate the deviant patterns o cunog in drva Hala, The average amounts of copper head, manganese, and and found in harta of patreats in the age range of the fourth through the ninth decada ere gien In TIDIT 1 Imounts are expressed in mg 100 gm of dned the Our urte i study does not embrace a sufficient number of hearts to establish poros for the ther melab. Adlounus f iron m these patients are much higher than the amounts reported m 1951 and ne are forced to conclude that the earlier nwirt as in error Ina ooperatne wody nom in progress in the pathology department and the (anwings Ketion of the Department of Velicine, Unlversity of Southern Cabalordia Schonl of Seccine e art sehing to determine whether conditions such as Diy xarjal infarction or atherosclerotk baart disease ne nalrrored in changes in oncentration of mundul tnce melal or in altered trace-acial patterns. Llegde young shaxman from Ina working in our seciloo, hat studied fifty bearts In the past ytas In auch in dre he has obuolned spect mens from the right veiskile the left ventricle and the interentricular key tom Where coronary esets have been involved he has also obulae sec 11ons of myocardiam mloding the invol ed arteries, and in serial Inxuncas he has taken met ons from the airta, the north, the enda ardlann or the saplary muscles problem coolronted us in determining shah arcus to ample In the Audy of heart true from patient with infarction of an area of the left ventrick for etemple wasit pecessary to wropte both the infarcted and the coalalarcted area or would the metal patterns of both areas Le policiently Hollar that only one sample would be representati e? le found that only the one umple was nerary unles both a new and an old Inlarrt sett jelent le undecal are in establshing our dlagrosa. ben the bearta WOTE gren to us by the Antony karton they love only in entory number ve had no clue as to the demon Hegole stened the sections in hxh uc The art mporte In the gorted to rent by mut from Larsele Lab aton, Camibe Orange Conty Hledt soruto unte in GW and the Su Perundmollart i slation, San Bernarum lub ·Gudith Heart Aſuscle Ion Concentration 391 Tan 1 Jum ra 100 G D TMT 1 71 N Loud SE N Mungen SL N Zinc SE N 1 42 03 4 0 256 0 066 7 0 116 0 017 7 12 00 105 7 18 0 24 10 0 212 0 0.36 11 0176 0 012 11 11 10 0 97 11 1 34 0 14 23 0 212 0 000 31 0 143 0 0 193 0 18 37 0216 0010 U 0 172 0 024 1 93 0 23 24 0 117 0 02 31 0 130 0 000 31 12 03 2 22 03 18 0 191 0 034 20 0 153 0002 20 12 80 1 70 20 11 12 019 12 17 078 4 J1 Areng SE -Aurd ETTO N-danber olara ere Interested, labeled them, uod establuhod the durnos hustologlaally Ho then tnced the parents chart through the autopsy number and checked his finding gunst (1) the clinical impression of the patients phyacan (2) entres on the chart itaell and (3) the findings of the autopsy rangton LD most instances an electrocardiogram mis available for confirmation of the dagnon. In aika of muspected myocardial infarction, . kum tamminano study wantly su avulable Patients were grouped us folions (1) acut myocardul infarction (2) acut and chronx myocardial infarction (3) chronic myocardul infarction (4) atherosclerotic bart direk (5) Ilopathic bypertrophy (6) endosclerous and (7) amyladoda lle have foot received pectrographic analyses of 13 metals in the fint 32 I 50 houts included in our glody but we have not had suffraent time to make a thorough analyms of the findiops and to check the clinlaul hustones of patients wbos metu patlerna deviate from tbe normal Honever we have made a pomber of interesting and greative fiddingh Thirteen metal vere stodied in these bearts land manganes, zinc, iron, copper molybdenum alcium aluminum cobalt alrer nichd chru.solum, uod admium Lend content ww cutremely low in three patients, all i wbom had greatly bypertrophled and fubby barts with thickened left ventrcle A single patient with lead martoly ekvated in his heart the proted to be en groot enlarged flight ventricular wall and hoge I rer A xrond nice metal ulied, mangane provel most roteresting When areas of recent infarction were analyzed the marginea con otration su foundt be quite low bat haureu contai ing okler Infarcts the coocootnatrons of the metal cre markedly elented As un illustration in one beurt It both in old und fresh Infarct the manganese concentration at the ate of fresh infarction mu only 0 03 m/100 gm of dned toue but la the ·392 Annals Ner lorl iademy of Saences the old infarct the manganese value kuu high us 0 29 mg (the 11ttage value for coapacte m 113 persons of ages between 31 and 90 1 0 148 mg) The concentrations of both anc and aluminum ete elevated in two patients *ith palmopan edema plus intcnual herborrbage and me third patient with intrcanul hemoorthugs, but without any greulon of pulmonary cdem Chromium and nickel cre lonerad and alt ru elevated sharply In a re tient with epilepay who had suffered a rupture of 20 aneurytor of the splenk unters and a lowered amount of cobalt was found in the heart of a patient with hemoptytos A 90-year-old woman with intordalous ongin of Ibe let circumfler artery markedly wchrased coronary artenes, and hea y calification of the mitnl and inauspad valves us found to be e 506 mk alaum 100 m of dred to in the tricuspul are und 388 mg akuum in the mitral val e ara (the range of calaom in the hearts of our 32 patents in tbe urrent study 5 8 to 160-30 mg the average value for the 3 meraires) Tor our initial Audio, representative values were btained by avenging the concentrations of the metal in the right ventrale the left ventricle and the Il reptum As we go int more detail me plan to give attention to differences in metal consentrations and patterm in each era of the heart We are now trying to determine why monc patient with auto and chronk myocardial mfunction, for cumpte absence of delectable molybdenum is worried with Dormal levels of the other trace Detals and in i krood patient absence of letectaWe molybdenum a companied by a boence of cobalt and chromium in the ventricles There are many oh situations in the investigatbon, whatever metal we are studying Inlerde orth Betamax 3Idals Compet Relationships of other true poetal to molybdenum serie to illustrate the interdigitation of the inic pells in the mutntion and metabolum of the bods Viord in tigators ha e been asure for some time of the antagonism beincrocopter n molt lukenun especially in animal naintion Copper le ficiency mualileo budueather to low copper concentration of pasture grures or to high mol lich um ontent However I third factor enter into the mtution, and perhapn fourth The degree of molybdenum upptendon of copyker depends on the mount of Inorganx allate in the dret and the com Luned inhlbuton eflat of the man vtalenum and the sulfate on copper otdination SOTTO I depend op on the ammont of manganese avabble lligh mangane ontent iti Hond the way bdenum-rullate surperson of copper storage and utlization lle maut all further relilxation if the diet very high in filein, manganese mis augment rather than lock the effects of molybdenum and solfur on nyrer atiliation Inother endte metal bat parently interacts with molybdenum is tungsten, khi Linto mhilt the u in in of nestinal senthine orxda c. Jld Internal Il ya lox Ian Shruover ha HTTU culating the rule of the trac melals in urtetaal byterenym Vier the disempty that hyunladne in ·Gnfith Heart Vuscle Ion Concentration 393 ADN pertoe mot Kelertney hinda manguere ferrous mon XT aver en and mencary and that by dostane potkrad ostb tbe aber tout of murupen by nedom and GTX won, der mestptos lented a trumber of chelating sod metal-bindmg gedte end found that men would love arterial hipertension. They then anal ud the KTOR & struktures of nt hypertenche molumns non * or undergoing dinaal trul, and found that many apporently doer gente, none of hxh act on the autonoma nan, had the comme property of bending thice metra Occally PETRTs trailed with simsporoasa (RALI * esbylexe damistemacetate (EDTA) tro chelating seats that have been wred to lower hpertention, respond pandacally th a further in blood MAST Toi muggers that mobilizato77 of metals as affect ranglar bordeonun in alber duetno Schweiler dele a base wolled pepitke, presentasin, from the wiera blood of hypertemuve paleta Phetentia in macunlod by ach of dr. metal-biedny a obelatung Acou, providing endence that the construction of Taxcular wooth movie may be loftxnced by a metalopepeleke the blood Cadmium, bend, and mappinek are found a max amoenu m the urme of persons with malignant bypertension Tho suggesus that bere metals may be prosent in hypertengre patients in abnormally large ASTOND u might ha e bore predicted when hypertensive packniS use tretied with you sucks as bydratazna, bigt urinary levels of these metab drop toward normal ET ureuon of orre metal, vendarzo, considerably less hyperlove parnu tan to nomal perons Only after a large Tro of cares has been thoroughly studied will it be ponddate to determine the role of metalbenuty'me mbalne L encrul bypested and t sportion with certainty ibe contnbution of euch truc metal to that ombulance. In our laboratory we shall be mterested in metal meya of cocke pics from the heart of hypertente porenu i our te Ndals * Adhood Sletals also may play a past it atbrauc KTO.lle huse entence that metab are interey mooled bis fatty acid metabol ror cumple, Car run has thors that an nu chromium cred by 150 per cent the incorpora shop of -labeled 10eute into colcnierod and atty acidu Janganese long recognised as lspotropbox agent incran kynthed by 123 per cent rana diom detta vorhers by 0 per cent The mochctxoo of the telator &-bydroryaingiac descasar chokered and fatty and tynibers, pagering that diferent metal ur mobihed or remoosed by the metal-burung agent nals in Renmeie Fas The detection of arc coproporphyrin is tbo arie facute rheumatk fener wiges but zo may be trained in this beart-grippling din Orien the rurkular uppelepas remoned routinely durtas mitral romanbuotomy thor Rims of urine rheumatk dbruk He have been maying that tissit and plan to was that once weial content by nem a the etio pecivo graph ·391 Annals New York Sademy of Sciences Udals in Congestire Faume We are laming more and more about the roles of ons other than sodrum in ibe pathogeners and control of congostne failure Jo the doc conducted at the Los Angeles County Hormal me hare mamtuned patients free from ugnifiant reoal dinLX for months or yan on modente ar mwyt doses of organomcncumlı sithout mjury to Lidocy function, provided naiber oliguria oor hyportena u present Analysis of the Lidney true from +5 paixents with congestie fature aboned an increux of mercury deposh on the day with continued admin natron, but are no endere of mterference <ith Ladnes uction la protein nitrogen alue for these paixents could not be interpreted u indatog that the larger amounts of mercury mtbe saves were harmful Lowel loonnus the method of action of carbonic anhydrox mhibitors Here, too a metal B of primary mportance, for arbook anhydruke mhibaton work by blocking the actma of one, the metal that is an mdspensable part of the carbonic anhydrae molecule Removal of the I rent in reversible Imactrabon of the metalloenzyme reversible macthabon us cffected by enzyme mhibutors that combudeth pinc, each as dmeraprol cyande sulfide, izde, and the pulfonamides I shall not doncu obe dies affecting the cardior exular system in which metab may play important roles, but I shall outline the paths our own iDNERE pato follow (1) He shall make thoroagh study of the metal concentntion in the beurts Dom under hudy and shall try to find an crplanation for abnormal metal Concentrations of unusual ince-metal patterns m the patienus duxal het tones. (2) Hegde aboo bus preserved additional theos from the surrous chambers of the fifty bearts und Hody There II be ruded for enzyme pat torm, which will be compared with the these metal profilen. (3) In mon matances the patients in our wo with acute or chroox myo arda infarctoo ba e bad serum inmammere studies m the period belore death Comparsons will be made between the sum namiek Aude ind uue coryme patterns (+) Be be e made udgements to moject nudboactre-taped mangrdere into patients with myocardul mfarct mom and thus to follon tbe incorporation of the tagged manganese mto kerum cholesterol (5) As I mentioned pretposty by means of emocion spectroscopy xe plan to study trace metals in the aurkcuber pendagas of petrons undergoing surgery for utml commneurotomy When we first undertook the study of metal coactatntions un cardoricular upd related to m the early 1950, IN COOriced that we had in the metals tool that could pomul u to dwoke and trat pathologal conditions long before these conditione bad udianced 10 the pofat bere they could be detected by mam of the slide and the MiToscope. Des dlopments have been even more npr than I envponed, and exdence is mounting that metals hold the Ley to un entirely per dunetLoon of medical matto that will coc day enable the du can to treat deux «th the accury aperted of the change or the physicist ·An da Griffith Heart Jfusce lon Concentration 395 { xoalegreat I than Edward I Bats and Balakrishna Hegde las cher coopertoa in bo sudy RefMan | Gum, OC F * BETT & Jun 1934. To norus herpent cortator corta bonne og ha tetonal Vodu 01 1 DAT XL BCL 1915 Soot preturnury Set on thet a Doh bomo soxtabolists betri o ursus hann J170 # B LB 19 per Cord fontaba WWW XXL WeCLS WOXT 1985 Cosges och beton arction, induction of by pack Vaart iT 171 5 Da, AT 108 rots bakerstra and the ration Sow Seat tr 279 Hos ED! Kaoh letett. 1956 Competan solo man noty lodem ponton Fedem Proc 774 7 Pun) itu.HSC 1954 Standao on 15t control hypertenson br Hyper In Pbermacolat od cokol obat alio 1 dracsopbichele Wed So & Samoa, UAE V kruum & H# par $ 1955 Lathyperten haud petal body was ) Cle Ved 4. 1! M N $ immun & PL SIM 1935 Fitts Abhype letrare not como mod decarboni alion and text oodation Fedo Frek 16113 10 Outll, B, COLD SI CALL 1946 Choral o o 21 decreapta propubel (I Plano donc obert SA BAL by more text JC Lara 43 11 BIL, SHKXK EXEZE 1932 The eet of doorcaprol os portas Lancet 2007 12 SOKKUR, SA S Olen 1950 Promo buenca utaal byporten Destmi opbermum, mots low procured from blood Expth Ved 4.55 1 Bar, H AHV Pant} E O Do&LE Want $955 Prawo Tel rypertension Cheron) and pwrmacopoul characteruta prestan Erpu led 2081319 14 Sox, RA 1956 Tract petulad dronc dana Luce In Ved 13 Comu,GL 1955 Voel dat gas and bala colonna be Prot x ETI Bu đỏ hi 10 18 Arx, OLC Von OP Hat 194 Lootrophw.on of a J BI Chen i 70 17 VAUX, BLEID ALTO CLIE 1949 Las the Top ta puterworrect to book uydrex My h 370 18 VALIK, BL 1956 Tb peaba toke K As We Are 18: 1053 ·VIAINTENANCE OF CONCENTRATION GRADIEVTS AND RECULATION OF CELL VOLUVE Alonnder Lauf Deportland UK Vechoros Cord late the Lord Imad Sabund and the lead Islas Hect law, Brion Veu It has long been seconued that a high surface-to-volume nto blasportant to rapidly metabolinng cells that are dependent on diffusion for supplies of orygen and nutrients from the purrounding meltum. It s also well known that the cell bus a high protein content istant proportion of which erlets In a soluble state in which it may be copected to crert an oncot Praire The interzutul fluid bething the cells, however a relatively low to its content of colloids From Audies with notopaally labeled tracers over the past twenty yaan it Dow well established ibat the cell membnse is peripable to water ud to all the solutes that contnbute significantly to the onmolality of the atracelular funds These volutos are largely the malts of sodium With these facts m mind one might expect that the oncotx premure crerted by tho intracellular proteins would aux dinerous nelling by drawing fluxd from the atracellular compartment into the celle Such swelling af cells obviously docs not occur m healthy toes in ino One therefore justified in Inquiring into the nature of the forces that normally countenact thos tendency toward selling of body cells and thereby preserve the normal intracellular ford volumo The maintenance of normal intracellular volume has been shown to be a procou requimg cperky In 1949 Stern ad obeerved that the selling of vzrou toen 1* turo wus dependent upon trouve reparation. They tested the effect of acrobxc veren nacTobuc incubalon In an motonic slide medium on the flud content of venous times of the guna pg. Whereas only amall changes in weight oa urred during herobic incubation, inaerobec conditions realted in a content large weight gain attributable to an inaakd water content of the tissue The type of oberration has been repeated by a numbe of other worken using various tissues and venous means of Inhibiting metabolum Aludge, Robinson, Deyrup and Whittam und Davier' have all repatedly conrmed the finding uong naerobiors, chilling dinltrophenol, unde und other metbods of interfering with tbove metabolism In luntancet mbibillon of Detabolen bus been found to be trocuted with card water content f the throue studed A much older observation that goes back at launt to Sabbatani s mort at the Kart this century is that meling of these in the can be prevented by Abattuting hypertonic solutions for the wul botonic allne media Gomon and Voor Opx Robinson, Acbl, und obers have confirmed the fooding The obaration that time selling dependent opon tasue metabolism The orl dexnbed in the poorpported a part by funds frog The John Tartton Foundatiog, In Na loch, N from the N to Nart Indt (Gnnt H 2X22) Punk llak kn Bethed, ud and from the Ameriu llant Amoontoo Talat NY 190 ·Lal Maintenance of Gradients 397 0 11 NO to Irg TO 76 74 0 QUL 04 00 au 10 to TORCITY PRO 1 The relation bets au cate of that when and conceolation of be moottor Detap Tent HR toplex of tara d Lab On the ONU thom ccto te of the bodo prepared as factor of 015 Where od on the ordinat the aber content of the (rat de carte de aprewed a per cut of the cupbt The action that perpeadicale braka sandala the more ater content of netbolung tu pedon The corre on the chung cocentro de hon for metabobo Docted a beta te per Curvt represents the ster content of bone metabolica anbed by natate loc and that it can be preveated by ertoox medu bave gi en met 1be com cpt but the matracellular funda bare a burber Oktob« prematre than that of the atacellular iad and that the opOUx gradient prieteni ad int# and m metabolining the HKD libro by an active extranon a water from the cell Such an actor alonon of water to maintain cel volume kerred to be a neces Ary requutervent as Hodot with notopically labeled water bare atached the permebihty of cell membranes to water The experbontoul facts upon wuch the concept has been establutbed tre ammenad m noun I wbkh xa constructed from dau of Robandon On the best in plotted concentnton oltbe medium ctpressed u factors of 0 15 1 mline and on the ordinate born the stat content of the ove (ru: Endocy cortar Hica) cpued us percentage of net te maghi The point of Intermection of the 2 perpendicular boken met dates ibe Dormal unter optent of metaboliang x n a mediom Bionx to trudlutar fuld The long curre ebon, the changes in water contrat culture from changes in con centnton of the pedrud fox metabolit time bcubated at WC bereas the oppet cunt represent the water content of the bore metabola nu phibited by excubaton at C Intubation d martabolum thout change in toarity of the medium s * to produce e larte te in base water ontent Orer a comederable nu of con talration, abe water contents found to be bigter heates metal ·398 Annals New York Academy of Sacoces TAN 1 CIAOOS Tot Soort x, POT TY, CALORIDE, AN WATD Corom or COTA PO ID#Cory SUTTON I 110T AT O C Slca d Time (011 02.) Incuheted 0 to 50 min in 10 al al Biarloost Saboe, pH 74 Guus Mere 0+ CO.95 5) E4A D) at Water TOT C (1) Belore la cubation (b) After abution b- 14 23 7 0 35 3 22 +004 350 + 18 772 + 7 365 8 14 19 + + O2 + 16 +003 579 + 18 417 + 17! 240 + 7 +0 94 0 0 +229 + 24'+145 + 18-125 11 Reproduced by pero o Te Duchenne en Van valaa #5 olismus inhubited than when the inpue mus metaboliang Furthermore, an IDCTC in concentration of the medium, in ths come to 1 65 tunes notons w effective in presenting the selling even tough metabom u in hibited. As rated, these findings have been interpreted as evidence for a hagber mincellular than estncellular fork procure that is maintined in metaboluan ede by the acthe optroon of water u npdly us it enters tbe cella This ugument s supported by the evidenco thus far presented nly 1 tbc the * dung due to net shifts of water done into the tous If det movtmeu al otrnornally actult sute 250 occur actosa cell membros doring ach 2* ifre studies, such an interpretation not justifad TALT 1 shows the effects on the water odru potulum, and cblonde content of 0 1 to 0.2 gm slices of guna pekxdney cortes ben incubated 30 to 50 min at OC mawlide medrum umilar to medium 3 of Krebel Without addition of tbe organic abstntes Control sacos were umincubated, freab sicos of the Results ut aproated per Lilogram of the molds. The Tunne selling s odkated by the locTonke in beste water 091 Liten. The incax in time sodium and decrease in true polemium noted by others i meen. Of perluaps more importance to the present argument is the large for Cake In the chloride that occurred. One an alculate the concentntlon of chloride in the increment of water med 151 mEq 1. As the concentration of chlorde in the exubating modum su 135 mEq/) the data frongly west that the pain in there water content in the experimenta reputxated or mentally an isotonic @try of modo mto the time. The inding forubdato the norimpion that ID melling is due only to net movements of water between taxes and media Smilar results have been obtained when other mamamlan to have been stodred and other man of Inhibiting there metabolism utilipdu There Ictulus confirm xadxs by Sludge who had prevlonly shown that the te saling produced by & variety of mhibitors of metabolism nu toated with an optake of love u well us of mater by tbe mallen tap. Our findings are ·Leaf Haintengnce of Gradients 399 fully in accord mith ho coactuswon that depression of metabol activity, Therefore wmawaidd when motx incat in cellular bydra Lio" The prin in 100 maler and salute is the present experiments probably whered by both bo man and attacellular phases of the the Thus, Robood mind by metrarement of the main and KKTOR uomines ant nu! corter at the atracellular Water remaim to approximately content fraction 20 per cent of the eight of the wee hether the hydration normal or increased Tou finding has been confirmed with #Kes of guida por Idaey cortex oxubated arobally al O C bollbsttam." From the above its apparent that the se selling that occur when retabo matamied results from a movement of both wluto and water to be imma Tha midates the aparent that it n due to water alone moving in response to an ook gadkot Thu type o endence annot box ever provide an answer to the foodamental question of bether umudent domu un lact exist betucen ibe water atently and the extruelolar fud and that of the fland within the cells To ADEX ET the question, ac must turn to a mone direct experimental approach lo 1901 Sabbmcam bought to determine the freening point of varu doktor et und adxusel values that everaged about 150 per cent pater than that for blood Collip Lorry and herpel-Fronica, Cacaosors and Sabar Patchotka a al al confined Sabbaiana' findings, udadt Open Dung melumak point depuis There ctyocopia mafurenenud have been uitxurd, bow crer because auto occurnog danng the sterna beirceo removal of the te from the Rural and the end port of the mawrenent will yuld puru ously high what to lieke condity Thxt raped area ofreenang poat dquthne of the mil octar er at Chu been demokrated by Conway And JIcCormack, Conway d d d by Brodsky do Conny and JcCormack claimed that the freezing point depression of the corrected for the empected autołyos, *** the same u for the xtum Brodda were unable to confirma tham ettipalsted vaha Honever by beating in IS in mic at 6°C for 30 min wedsttely upon remporait pedece xotol yms and then paming ibe freening point of the besied nutart, Appelboom has recently reported wolonialy with tracellular fukis la our laboratory LoRoy H Jaliy metly ha atulired bc* echnique for obcamisgameting curve Muals wete fruten by me m liquid artrogen medaily opon removed from abortbriived a pumata. At the scraperature of lad ostrogen the towe nu pulverited and ibe finely Pound powder spended unett, chilled, bgaad alicone Thu yuckled a lourd seen of twee that could be started vigurowly and warmed mdoully in the premise of a warnt teroperture mecany dent. By the mean, tx autołyu kus konded, unce therruul gradients the tharme Delub of the wody <ul moon be published dren here but noin 2 Home the molts al hus obtained for aduc le of ihe rat Here aro ported for ampamonike midal portion of the melting curve oni bart normal at rom having sennion of 311 mm (u manual by so exhepensioni prece i cering point metbod) kom tootaunted the hydratko o 50% mom and Krut dilated to 100 mes ? Tbc ordinate ·400 Annals New York Academy of Sciences OLMU 10 / DUR pod பயம் TO 0 0 10 10 40 00 CO 70 FOTOMTOMETER FIOL 2 Song cunes for at heart ( autoler brusche) courpared nth thoma normal nt om iad coactarted ud tobuted at wrum Biorements art nude on portions or wrum containing spul total quantities of ta Tbe and at the perconut oths ter that bu mehed and the same the potes boneta rodong back proportout temperature of totul plut concentration The bonupaul dostanet be toto the cuOM mentre of the clarace in hi! Cuocrotnoton or a water activity between the ripectn curva Excurve sprements the meu a 12 mensarments and the well rohallinor ros o dous thon the mixtard armor of the ma for ad poent plotted It accent that that u cheference between the monuments for baut nude and for the percenug of Ke, in true or kerum, that has melted and the bacima the potentiometer Kule proportional to the temperature of the melting an pertron or to its oomolality Each curve the mean of 12 separate marre ment. The standard error of the man for aach pont shown by the small vertal bar or tow of dots. It is evident that the values for bart murcle do not differ ugoufiantly from those of the sun Sunilar renults he e been obtained by Sally for skeletal muscle, liver bruto, and kidney corta of the mut These direct marremenu prove conclusively that the materactity within the cells u the mme us that of the atacellular funds and that do not gradient arbus between the intracellular compartment and the Internital ftud. What then, the force that au timpus to smell ben the metab otisas ibibited and how thandling proiented : ine and in metaboliung theses In turo? I propose une vien that consistent with current koorlede of yon concentrations in ta It has long been mognued that the fluid contaloed within calls La bib in potasiun and lon in sodium and chlonde concentntions while in the estra cellular turds, the relatwonshup is thered that is, sodium and chiloride loms are present as the major soluta while potanrum exists at low concen ·Laal Maintenance of Gradients 101 intron Sirxe sode with anger hose momentet tut the cell rock brana tre permabk to odmm, potunus, chlork, and matet kodeot luat muintenance of the roo gradients be tongues requer the expenditure of Spedilu kthe tampont netus bure been described far modrom * poiam, ** and chlorches su cells and traie Whether there transport proces paute ungly or together in all as to extrude codrum and hlaride and to accumulate polemiamo ordert mamiana the characteristic non gaudeats o act as bng moddertered The obtiene mudents mo man moet coolt amply itauli from an active outward minion of pod um alone from the cella, aith other om faller ung pariets * Recen radies doic that the actual diaction is most compler than the und FTER AD mts runt process for at least polnun us wel w rodam. In ad Terca, honeter cells utilize niet dertied fro metabolism to og Tule ther to pump The interior of sont cells electricalh nepune relaine to the exterior of the cell The clecimal polarity of the cell membrane tostated to the content on the col of bred or nondiffusible asjana group on matracellula pro tems and other large molecule and to the outuurd ertsog od rodam effect of the electric potestasi croce alle a to contribute to the concen I ctbande and other bonuse charged wall wons of the medium The ninacellular seguim electrometrally opposes the ice negatne charge f harde ku Then meubotom phibsted, the energy required for the won transport prontu Do kager i uke Sodium that duftuar como oricirim mudent into the cell an na koger be entruded poumum land out of the cella tato the murrounding rutrucelalar fuld As a mult of the acumulaton of sodam Low cell add bou oli potassum, the metabrane potraint o redu ed to that chloride u well as txtum enim the cell The rola mu gretter Altry of sodiumo than low of polisrom by the lame e thon TULE 1 The net pris of solute by the INC succompanied to an BOROok gam Austes and the rate becodes solent The procesin tuable for a time batut-o thut rampion of metabolumn curs the high flord ontent of the nollen tatue to retus tonurd parmat Laille demontated 1907that the rata premere certed to a men on Muth of proleta detox with mask ourentnies of neutral is in the medium It be theoretxully posible bendore that ording of LOKAK ay be presented me ben tretabolismb beled by pung Chees m bypertonn woh mlavou Thai us thorp in Mou. In exactly at Open Robinson, and delu have found eyeball Ji catr from hat has been stated abone bonne that ths finding to fit the casm that for port urva troba a the othptery oy of blonde wito di te Coro dictabo a patibetad, so Lal * Ad card permeability to ali so to wydatne is the on celet cention of the 100 bitabolated Wowodus oodato are the codi andato a thu satud entre of the own to pletely wbucal Asyet, som har start the teton bich cord hoobin permeabbaty te permula padengthes that woultrabok + } ·402 Annals New York Academy of Saences bo 在Xy No No ar No or AT Fote 3 Scentc roprementation of Dormally pretaboludok el (add cell home metabolum has been tembly inbiboted (b) 10 Borate the important factor rartikel cell vobant (A) I the normy Detabolang Na dudum into tbc od odoot, us how on its roht hand woke However tryt xoa tort indicated by the rotating band on the left Duran ol the cell, atruda N uod and Kuptate ocent to mental high oth cholar K contatud low cooker Na content, respectively, be the Rady wat тъ cell membran potou a thon +) onde apd (-) mode The probro poteptlal depicted u preventing entry of chlonde into the cell Anorerents conducten ble poly malat trzebuer wrons (such as proteine und nochec conde) (b) The atonton a ollen celo depicted Inblution of metabolum thor or stop them pump to tbut tbc N aby the cell by dios annot be extruded and the bout from the cell Cuonot be replaced The cdl nero brade poteabel dominube and cond Dow at the cell The requres further mtry a N hat the cell to pra ckttnc Drunky Thomard plut catal of the cell reo n atoodan miry of stered tho cd becoo oo Whether caso in ce men brade per ability to protry of 1004 to occur ben petabolibaba bited and contribut i the catry of solat lnt the cluye unkon the bich concentration of such media represents the true total solute copcen tration bio metaboluong cella FOURE 3 rummarpras schema lically the ven presented hert. rigou. 3.2 shorn a normally metabolining cell or wual volume The content of pondiffusible into cellular maternal, largely protan with pet negative charge at physiologial pe, ls indicated by AE An active transport Dechanika u depicted as a wheel routing ma chochote direction to etrude sodium frorn the cell add to take up potratum from the median at ante suficient to main tun stady-tate concentntions of these small lom in splte of slow put duron of sodium into the cell and lalege of Inncdular potanud to the medium. The relative concertations of kons are roughly rodeated by the dre of the letters The oncotte premture Ithin the cell la thus ofiset by the obligatory atracellular position of sodium, wbica la maintained at the empenre of may from toe metabolism A double Donnan tysten bs met up, and swelling does not occur ·Loaf Blaintenance of Gradients 403 fronu 3b shomthe effect of mhibuting metabolum The unty of the on pura la reduced that I can no longer estride sodium or take up polar hum u napadly as sodium Jima dovo its concentruoa grudkut into the cell or as pourmet from the c! The result kenet quo of not cally utist tolule Ithin the d und the idrok A retur of metalonul result in sodnim einson and pornm uptake ith return to the unwollen normal suale. Finally I make me further fpeculation What the raficance of the mechanism to the cel? The alternative adjarmeni powible to the cell ca prevent the wellung that soukrout from its content of intracelular collodoald be 10 Aurround itachi Hith a nu cung 10 withstand the force Honnet high membrane teraron would be very daadvatageous to fall cells, which are dependeat for many vital procenter on a high surface volume relationship Furthermorem those instances in whoxcb membrane tenors danmal celb have been menned, very kom wahes have been obtained. E Harity cammanned tbu finding by rating We are satisoned to thinking m tams of un-rile erface tem Son of 73 dynas, CTO o that it hard 10 realur that the term of cellar keast 1000 lmes perhaps 10,000 topolom. There lo metabruor tenant are pouble I believe because the anum-ul cells have learned to avond dastrous wellme by developing tbe kom pumping mechanam moi douane (crtad cela sem to have cvohned secondary is for the merky stored in the wa grudi ents or in the terulung cell membrane potentas Verrou ooductivity kod dokular contractity pertapa, represent aimples of such condary evolu tonary adaptations. Thus we one the miraculty of the myocardiam eod, in fact, our very mobility a rozpared with the suke member of the co 1able kingdom, to the method of preacing cell volume wbkha anche the cumbersome cellulose cun that the plant cells chore u thor na to be thurert of cell swelling Refore. 15 JXLI EorumOU R Meow H. Kun 14.99 acbeo J4 410 Icon, OH 1951 Aj Phyto 1 06 Roman B, 1190 proc Mary Soc. Looder BL17:37 An ) Phyu 17 39 3 WROTTAX, IL DY 1953 Boce 18180 Sunaton, 101 Adet path $19 7 Goeder, PS Haut 9n Arch u Putler Phenmukol Nuor Sc be 1 149 OF, EL 149 Egu Wed 183 9 ADK 1 1980 Her Pop Arts 19 10 KADHA 1150 Procha n Beoplay Acta 240 11 LAAT A 1956 Darbe Du 24 12 TAK 199 m 101: 441 It hea Hon that med may suy acou o poreon from the Lody S dan could not uued minskar omok pour slut och The tota u TALE 1 bonne whout that door etter okat ere the datto Kate the powe thold the Died for ury dcbo que el cate us the time Forthport the procedure di comed e trouble Tor these me the powebke tor contribution of motore the preko daudbe the patte, but would น ·404 Annals New York Academy of Sciences 13 C ILIJD 1920 J Biol Chem 221 14 LZO E F&E Kan I nos 1931 Ireba mpu htbox I'haurdulal Ano - Schmiedebery 10 236 15 PTTEPOTEA, JH Horn & J RDS 1954 Arch apt) Ptbol. Pharma Nanyo Schmiedebar 311 16 OrL, EL 1954 Lopullie 17 Cox LJIJI CHL 1953 J by w 110-1 16 LOT E) I Cano Ji Weconci 1955 J Phymol 180: 417 19 B ODUKT HAJAT.001, 1 H DINS ROY J S JILEY & I DVD 1956 I Con Pro 4 183 20 4 1957 Tederation Proc 161 778 21 USD, H I &K Znan 1951 hete Phymol Sud : 110 2 Rot ALH Los 1916 Į Celular Comp Phy wil o 231 23 SAXA, MULD BE 1953 ) Cellular Comp by U177 24 Ku, HA R Watt & Ross 1957 Brocbem) 86 53 25 Hoon CAM 195.5 I Electrolyton Brologal System 176 AH Sbude, Ed An Playzo Soc Ambngton, DC 26 STOC HB 199 173 ESG BUTO, Ed Aademic Press New York NY 27 G UDIWT 1955 7 Electrochemistry in Badoo and Medicino 147 т Shedlovky Ed Wilay and Sons New York, NY 18 SEA FH8CSwor, BI JODLETONT & JIE HOLAT 1956 J Geo Phyno 40-263 29 MUDOL, G H 1953 AD J Phymol 173: S11 30 SEANO, A 1955 I Electrolyta in Blodopal System + 157 A Shane, Ed Am Phymol soc Hashington, DC 31 ROMAX, AS 1956 Yake J Bal h 218 32 Tomc,L Y EOULLITOY & R A Em 1050 Brochet J 47 130 13 KM, HAL V FOOLISTON & C Tam 1951 Biocom 48 530 31 MIUDOL, GH 1951 Am J Phymal 1 113 35 LILLIL, RS 1907 Am J Phyl 301 127 36 HARVEY EN Protopheumatologue I LS 1954 ·PHI SIOLOGICAL EFFECT OF IONS Rachand R Overton lotte (Cluxe Txerit latersty Tam Costs of Ixis Nemper Tan Ixiram At one mage in the development of our modero concepu or renal function * nelloons phytologin mand, Anc Loon fw me about the extra natur * sals unse* Although he literature abounds ush kristniau theory and derer patrunking and even thoughtful experientation regering the effecta of kom on curdac function, one khely to conduce upons perisal of the ma letal that I wehren er syns about the play wokgralchiru of us that there are some It should be added, lo haste, that following the quoted at terance on docy lunction caucho las been laimedestigt neked, brus on the equally buron remark that be Ladseys the only link torpad in the body Although are shall see, oor present poatxos repardong von effects o prednou, wc thus bare micellent redne to bope thar umiar progres will occur ta relation to tbb problema Reviews bank deposition, ex certain (boib ketual relative) *od arc morreret, eod thoone of waic etfecue Dede izrety ARTE forms and es abound Ilich abse combined adrent of Holopac and flame protometrk method, the fourth reports in by field are mapally m creuung in both mumber and rule Thus, ibe task of rette ny even lmtal porton of the field o marinental Cardio D posts and entrare Paen als The mincetobar plass of bent onescle dhe most emal cells, bus a highet K and lower la od CH concentration than u promet ma the extracellular fhund, bertus a appeun in apprormately eguralent om made And ootade the cell Tbc functional burner shat provides for these darbo cu nequalia . probably at estatomal cell border bat at subcutxular baya of the cell protoplam the properties of bich appear to differ from those of the contactous celt mterror The importance of this luuting or purtally Immunios membrane medecimy ponxe parton during boub set and activity mula berxel densiption of its probable aracture germane Iturred trore cumparatne Itar diüntido Audies, from water solbuty mantients, and from rortart Imam consuler sitors itaifa laser ol pre molecules produbly ovetisen som poudal material the combued them being of the other of 200 ta Gucisku Uhmiera the armate atracture borove the rating potential certainty related to the difference on pax concentraton on the tko skla al tbe fare troul barris Proceedmg from the potentional difference actuut the restiny cudisc fiber marotabe ( 0 to 100 ) and from batne Inow of aD AT mone reprdrag the Korek cox esintion of the mincelular pat staan loker that the membrane more percobic to ty than anunt Summilarly it 105 ·406 Annals New York Academy of Sciences can be seeo that "hule K+ and ct more still the potental gndent (bat in opposite dirt toD) Art laving the fiber must be transported ego in both 1 COD entrat on and on eletrgadient Vost of the statements usually made regarding ton disposition und more ment are only pertually true and we bured upon ret result of movements both directions the dynams of which we only wightly understood Never thelen, it has been necesary to post won theory us to the mechanion whereby an on an be innsponed against an chet trahemaal gradient, and its recog. nized that in the final analys Rich "prropog mechanion must depend upon cDergy ylelding metabolxc events sithin the cell Such theories may be nece men to explain mor of Koons unel as the election of Nalone, although Do fim evidenc for the former (u fur u the hart is concerned) bu yet been pro duced Assuming the foregoing to be approximately true, its clar that the impost tion of eternal current flow (action current) or any effective reduction in the metabolic activities governing membrane permeablity would repult in lowering tho rasting potentul to the point of allowing a sable na milers Whether Coocomunt depression in i espulsion occurs o not known but in any aume, the ortruson mecbansen for en temporanly overcome and the polenty u reverxed Thus the making phase of the cardiac action potental appens to be the result of the entruxce of Na ons in the cell The Calling phase, on the other bæod, w apparently the result of (1) recitry of effectiveness of the Na citruson mechanum (or reduction m Na pertability) und (2) an increase in membrane permanbalny to K bringing about a set out how of positive charge and tending to restore the resting potential Honever the pochopeni moc mied with repolanation of arduc membranes are les well atabluhed than thohe Lccounting for depolcriation The fact that the resting potential a not re-atablohed for about 200 to 300 meet indiata that in heart muscle ather the outflow of K+ delayed or the alteration of the membrane toxud again actively citruding Nat a vow relative to the conditions found in netic It has been suggested that part of the recovery phax due to metabolk action of the mitochondrn, the membrane properties of which seem to be such that Me can be held against a coocentation gredient Temperature Efes Eledne fials and Toxic Dus pasillos Trutnende ind, more recently Vanball and others have shown quite condstent changes in the membrano polentub of bolatod atrial fiber rubfected t cold In real, these changes como a reduction in the resting potential the most stnkng reduction appen at temperatura belor 210 Cung but definite down and trends are shown cren at 25°C The results of cooling have been to comment indeed that there has been considerablo gerulatlon that much polentul redu tons are the result of cold depression of the metabolk phenomene rasponable for ather Nat outlor and/or K+ infor Conclunya evidence of tbex rumel oor eventi tbo ftolated fiber preparations not available, boweier Studes on the effels of cooling lbe abole animal on ardiac ion dariballon have been made. u Sone of these investigations utilize coronary artetto ·Overman Physiological Effect of lons 407 bor Der 10) od To Too too so 401 VO 100! tol HO! roo TT SCO TOME 1 Avery lett rencabar Na, K, Cl, und HD por ky xet tud tbe X/N rubo for 30 DOTDharma dop, ter mnd cooked to 25for 2 houn, and 10 dog hanted t 41 5C for 1 hour The actoard Dumbent beneath the bar ut i valocs for the compen toos ondanted at the top and we al tbc ban Tbe bobifce naber der mat Kantials mukaat duderence, 2009 venous differences (without knowing coronary flow) in order to LITTVC t pomiblo oc probable changes in heart on dopodtion in hypotherm Olhe have em ployed the rubilied incer technique bat have often neglected to measure total treue lans Few groupe bave maintained animals at bypothermk heel for perod afficiently long to have been studying a stady-state mtution Recently Sparr d al" bave maintawed animals at 25 C for a 2-bour perod par to analysis of ventnxular murcle for Me, K, C and water At the time of uring ardac lance for adalyos, these animals bad realed fet clectrocardrogrupbxc abnormal the other than a mured ST wegment A portion of the realu obtuned i bown n MOUR 1 It will be seen that contrary to e pectatoos ribed by studia of clectic reporters of cooled angle fiberi add, in general contrary to other studies of the abode animal in huch nudah nere cooled to about 25 C aod umedutely umpked the results of thu stody to dante wafant reduction in ventncular masco Na od admibaunt nem There are so cbangu poled in al concentration norm water content of the muscle Where the angle fiber studies are excellent presentation of clectnc events and medbrinde potential witboat accompanying lonic date, the bote animal Hodies the far-revealing us they are of chemsal tony and even of boo dynamo-bare failed t prezent tumultaneou electric patterna ether a sati charges or activity potentiale Medmann has previously polated out the deurbel ty of obtuning data that allow dostinguishing between tunnen er in the renting beurt add the increment or decement due to act nity It probably equally important to male studios in the Heady state to uke ·408 Annals New York Academy of Sciences able account of pll alterations, to study multiple lon mon coedus all in the More prerantfon, and to attempt to correlate such finding with electnc a ents ar at least, with polentul differences. The purcity of information regarding for temperature effects on lon det bution in the beart is exceeded only by that relating hypertherma to bok movement and clectric activity TICURE 1 iodata that in the steady hyper thermx state (41.3 C for one hour) intricukr muscle pain no lat bat shows a K+ time that while ler than that of the hypothermx beast ytl gudantly different from control whues. Agato bon ever intimate Loon! cdge of rumuluneous electnc events (other than external EKG) is lacking Charger Concentration of "Extracelular" Ions It b generally conceded that increanng the ertracellular K concentration produces a shortening and a reduction in magnitude of cardiac action potentual, as well as a decreux in the microbrane potent Perfason with a K free solution on the other band, 1D C Car the renting potential unell as the dun Lion of the action potential Wedmann has shown that, in the polated turtle entrale, increasing the K concentntson in the perfusute cuns tramunt depolanation of the resting fiber abereas pch an coincellaber idctement ol K produced an curly and incomplete repolenatron of the Ame fibers - ben stimulated The blue of the resting potential then, 1 humed to depend upon the ratio of nucellular 6 to extnicellular K Honever the repolans ing effect of Kin the active preparation must be explained either on the bush that an Dcrease in outward K current has ocurred or that there has been a decreix in inwand a current, or that wouchou the one in extrellalar Khu sthmulated the \a pump and thus increased Ma estrusion Whereus the mfluence of Klunited pomenly to the cablohment or maintenance of a given membrane potential and to the repolanation procen ud an activt fiber Ne pripapally concerned in the depolanation proces Thus, in CTERADE extracellular Ne concentration leads to a higher action potential and a gente traboot "hyponatrenia" brug about the reserme. Inolar u Li aun replace Aa (up to appro tlmutely one third of tho esterol concentration) it acts as doen la In higher concentrations the effects of Li are more like tbore of decaused ertracellular Ne concentration except that it shorters the durition of the action potentul. The papery effects of Co (which are understood) are those of ultering the threbold and of antagonizing or reventing the changes brought about by K. Egal of"Jornal Inotropic Agents a Cai « lox Portin Recently Barn' reviewed the pandorical action of acetyktoline on atrial Limpie und related there pbenomena to lontc digribation and to changes in membrune permobility to one The normal Inhibitory action of acetylcholine ed its effect os incending the polariation of the atrul cell art well known. Il atnal ter cooled to somewhat loa than 20 C contrartions cause and actron potential are no longer propagated in the peripheral atrial time Under these conditions, however Aall, rythmk, bonpropagated potentbals ·Overman Physiological Effect of lons are still present in the pacemaker and, if ucetylcholue is added (uta beid delay) reguhr Dom impubos agun pear in tbe peripheral atrui mukul turc, and contractors are regined if the percerauker mpubes ubo drappeur acetykholues without effect ind, therefore, it annot be acting by depolalong the membrana. There finding hart been confirmed and attended by Jlar Kall Similarly if the atrla ve ullorad to bat untu Kby lovers the resting potential to the point where perpben/ atrul action potental propegation to Powupported, the addition of acetykholle restarta coatnection. It an be bom s also that nogal suuhailon acts in clctly the Amt way uod, further that the codutory action of the vazus under these cooditions aun be blocked by utropene. As previously stated, the action of acetylcholme Dot one of fiber depolan ution, but rather of amlion of anal cell permanblity to work, thus nuo 10%, the resting potentul Hoffman ud Suding he exbomo that acetył. choline accelenta repolanation matrial e end, u Kn largely emociated with the repolanation process, the would neemt be additional endect of the fact that acetylcholne action affects fiber permability t K Even in almal mock, which betrely free of real influence acetylcholine s farmoed by the colonc uctybas ryutem Burms believe that th: constantly formed acetylcholme effects in muitable resting membruse potental by affecting meinbrune perdability to K, and be hus shown that at that hert (turedt beat have lost most of the ability to form acetykhohine. If during acetyldrokino perfrisce, the atrial runte a decenat sumulated, Gibnlapon occun and will contime if the concentration amultaneously reduced in the bath Such fibrblation will perust unka (1) the acetylcholic perfusion & stopped or (2) the concentration in the medium rund Hoff mao and Sucklmgbr e shown that acetylcholine lengthens the period during which the fibers are exatable by wortening the repolarization procen electne dimulation can be thought of a loving the fiber out of phare then the presence of acetylatoline ppuently ladat fiber contindowly and werally stumulating one another and thus Grating In auch a Rtuation, remove! I cetykborne by loring the repourtation procent lengthen ngaun aka continuous auto-te-calation imposable The detailed mode of operation of the well-known epinephrine-ncetylcholine antagoorm has been studied by Web and Hollanderbo have sborn that pinephrine protructs atral repoination Of course, if the struc, then puxephone should antagonize the action of scetylcholme la producing condbon upder hreb fibrillation will occur Indeed, it huu pon been shown that electric poimulation that will produce fibrillation to the presence of acetyl coline la Ithout much effect of dreogfinncetylcholine mutuu ue etoployed m the perfuate It has been mentioned preponly that fibrillation can be prodced by elec tri xudulatko duno ucetykbolne mfupon only when the concentraton u redora Schubnllation can be stopped elther by TETOVOX acetylcholoon or by rarang the conceointion of the medium Thus kod's credence to x* that reduction in the bath of the action polearb} by uctyll accompund by an increased out and fur of through membrane ·410 Annals New York Academy of Sciences more permeable Such a view hu been strengthened further by the ndro Lotopx work of Harns and Hutto who bave shorn that a two to threefold Increase in the nate of low of Kº from previously Keequilibrated preparation 1 awoned by the addition of acetyk holine Their findings ertend unell, 10 kº uptake, Ithough the magoltude of komnard flos is comiderably le afated by acetylcholine than Ko outflus Their work also adds to the moont mg eviderke ibat the pacemaker region is chacutochtokelly differeat from the peripheral atrual musculature in that acetylcbolide effects the pacemaker and in its Kº permability more than it does the rest of the altrum Unfortunately there is little evidence beaning upon the question of whether membrune alterutxon brings about functional cardine changes through specific cffa us on son concenuation differences, inside this outade, or whether there effects are, ingend meduted through nta o change of bon concentration or en en porably by nata of penge through the membrane fa quotion. Burnl mata that varying the rate of electric stimulation of the atrta, in the abence of added acetykholide, from frequency of 200/min 1o 1200/min increased the rate of Klous but little (0 76 ml/kg/min. at 200 to 0.94 m3I/kg/min at 1200) On the other hand addition of ucetylcholine increased the nate of K lows at all frequencies, the rate i 200 min being 1 36 mI/kg/min (or about double the nate seen in the absence of added acetylcholine) und mung to a value of 2 10 mai 1g /min at a stimulation frequency of 800/min at hich point fibrillation enned Such findings tend to suggest that it is the rale of lor of K that important and thus presumably but not necemarily the rate of ud low Sed f Foreign Twaironic Agonis m Cordiklon Parturion Quindio acts strongly us an antagonnt to acetylcholine in arda mule, u well as in 10:00b und al cletal muscle. In the atnam thu materul reducta both the amplitude and rate of contraction and, if the export to provent concentrations of quinidine u prolonged, atral contractions cause Il after qumdine depression, acetylcholine added to such preparation, contraction begin again Acceptance of the mechanism of acetylchoke action as belo that of increang motobrade permeability to K ons land, then, to the as pumption that quinidine acts by reducing K outflus Quinidine induced atrial utest can be reverred by reducing the external concentration. This procedure increases the folde-to-outs de Kudent and excha kumlu to K effus reyena quinidino uction to fact tends to apport the umption that quindme hus opposte effects on membrupe properties from thore of acetylcholine. The analogy u not stxctly ime in that quinidine has no apparent effect on the reating potential of the membrane, hereus acetylcholine does Procaine and other surlace inesthetki and diphenylhydramine may act in Anber Amilar to qwn line at laust they all DxTease Loth the duration of the action potental and the duration of depolariation Digitals, loloacetate carbucbol ind phypostigmine ble producing no effects on the roagoltade of either the resting potential of a tion potential, act to shorten acuon potental duntion. Edence from the sort of Glynn und Schalmann, bo Rudied the action of stropbanthin kallaren and other ardic dycooda, lodiato ·Overman Physiological Effect of lons 411 that these obstacos probably cost their effects by altcrmgat ber nk perut tion or exc Damboused mmo Tuscapillor loric Toneros doctoon of on mobility 10 hat hus the appare of being a ampler membrane, the apillary endothelrum, peruncat. Although not much us Loom about it it will be apparent that our OOTDT of it at lart grat as our inportance of the phymologal effects of out on the beart) it will lastrate the long way we have yet to go before on effects art thoroughly understood There Do Coandenble history on theme of the sotope inter techngoc mrahng mformation concerning the druppence of wood from the ocular tyrem. * Although certain of the methods an be adapted to single to, most petbocs reported us uullised on the otact animal taal won a palery er bange rata, the remits being an alle brux Rim of muoy individual riteol exchange The methods such as that of Sheppard ddproduct data that mal a bort of vrables, they ha advantage in terms of emplaty and re prodoghulity and in pronding a brge amount of pertinent mallary data FIOTR 2 illustrates Rmultaneous time-concranton con for proteinbound dye (T 1824) \ und Kºfolomong the modden Injection of a soal Alay of these material mi large ein, the concearatom ha ang been determined m arterul blood obtamed by naple serial zpłmy From rach CUTTOS tl poble to akulat (1) the ardar output, (2) the model ardo palmar) coculation tme, (3) the central blood dłube, (+) the rate of loss al potope from the vucular ryttem, (5) the blood volume (6) the potope space udd, of men blood pressures aulable, (7) the total peripheral redouance. 102 18 thistedotat to do ti sto det & she who NOO For 2 The concertoo ou for T1R4 or montot atnipolut diled nakutarbee output ·412 Andals New York Academy of Sciences U n PE N IN WHEH N FROUX J Typical dumppendCE CLIVE odcatrog the late sono atrolat too The Lected dumun represents the kuhantom plot of the early pero porbon of the deepenace curre corrected by the vutul concentrations can red from the athpolat Inspection of these arves and the type curve shann in TOUR 3 indicates that, due to the heterogeneity of duppannce nio and the onset of lonac backflow from extravaxular poob encountered experiments with both rapidly o alonly excbanging rotopen, the curves do not remain linear and must be resolved into componen's Slope analyms of the arly operiodic portion of the Hotope curve (non replotted in the ineert of Tour 3) yells the man rate of change of botope between vascular and extravakuler pools For stady statc conditions, this acquel to the outnud transport nate opteed u the fraction of total Na or K tbat la as the arculation per muntle, c. vulve that llalker and Wikle aprewed us tumovers per muute Burlon a al have reported the repultu of such magrements in 65 normal dogs. Thox results indicate that Na kod Klave the rucular tyhed at fodividually constant rates, but at wynificantly different rata for the two OO peren Thus, the average yuxular stope (transport nale) for Men 1.38 + 0,30 while that for Ka 222 +039 The difference between these numbers bagnifiant at the 01 per cent level The doppannco nata for Na and K to highly reproducible not only in the xnx IDUDA!, but also from one animal to apother upd, Indeed from pena topecia Furthermore, there ntc constunts uppar to reflect fundamental properties of the endothelal barter or the Interaction of the roms with the baner ancestre altations in hemodynamo to alto the natador Endence has been presented by Sheppard dolm that the indivdultie transport nata do pot differ markedly from the man nate. Thex antbors showed that slope of the curly 1periodic portion of ·upon K Overman Phytological Effect of lang 413 Holope time-concentratna curva demed Ertra umpleng duderent seins were quite fra lur even iboogh Euch cures et various doplaced horizontally in tone and vertkaus in concentrum Theme nies, then, ire pot circulatron timated USCE # ippan ſom pais monog prom alterations in hemodynamics that us kons u capullana are avulable to the injected rotope the nie of bon wentical It alw appar tut all aplerse hart aprrt malets the one propria ufar u li and Innier in concerned Depute the duhealty Coconutered a changing the reported rute SADU they o be changed ma pomber of negi that prekat interung an enges of speculation regarding the lasctional anatomy of the esdotbelal amet Ex pomure of animal to whole body Wodation Ideas both a aud Tip port nates Inini con hutamine DCITREO Ma tunapart, but has no chat On the other hand, mentre cously admimatted hyalurondex - ICH K turnover nice about 60 per cent without ultering a trumport From the forceone it a clear that is and K normally leare the risular tyttem by roase a mechannu that are Bot only duferent burtlut an be independently or mutually altered hpplication of this method + labeled matchbox dx ferent moderulas sues and configurations bas au brooght to light sumber of mteresing and challenging facta The pucular skope (dappaurance rate) of natura radom, for comple, s about equal to the ruta tom of the alon the other hand, bench wohn a rubjected to cenatorution with tkali wibough there s bat liteke change it molecular weghe, che disap petrance rate a tery slow and approaches that of the protein-boand dycy T 1821 Eodacon of the wedu perenal of the capulary buLITET bed, inut the apertamental dekription of the rate of buddiedy other materials as well as the chcrta of other outstaro (moch u aldosteronc carda glycoaches, VILA mma) and peacia (bert, coll, anoma) upon these personaxeta Even then, only batang 1 hare been made, for Inom ledge of the functional capacite of this and other membrana may explam outh of Dewportines damos and rherapeuta As an crutopke of the combustion of the nute of tom masurement th tonal A ko determina 100 kod akulation of two and plagne gent actite and relativo specific ctions in the studimo font innsport and darbo ion canciers, the recent work of Hebeb klub Follow inchoar period of virus puas K-free Oure, the nt of low of K from the blood or dog nu determined, uel as the electrolyte and mia content of heart INTT and superal make поба жури м olent unduc munche su found and Lytt \mu bo agasanily ented The trean nat of four of K fross the ruler prom docier the condione (for example, 20 mom after contra o dulys) *u Wimbcantly reduced Such petuenta penerally rate more questions than they apnet and yet they warnt wportsat points relatne to wax payeukgy The RAI 10 the dearzbulity of capiling daix op i be MWMI O quatertuis inapon be send u abyrol Domace o naritan then llount the need for thes, i only of the stawy state of the wil of Tarrus punct of the angerous cha tomo of body Fat they beckon w with the wen ·1951 1952 Phyla 414 Annals New York Academy of Sciences entacing possibility that the solution of the problems will bring 1 ver a in the therapy for uod prevention of disease Roorences 1 HOOT 1957 Tbe I lectrop by wolor of the Hart Am NY Aad Sa ( 724 2 SATI, 4 11 (Ed) 1955 Llectrolytam Bolopal Systems Ao Phymol Soc. Hwhington, DC 3 Cun Hf (d) Ion Trumport Acrow Hearden Laudere Pro Nor Tork, NY 4 CLOTURY T (Cd) 1955 Mectrochemistry in Biako od Medictor Miley and Son Nox TorkNY 5 Mummy, Q R (Ed) 1957 Metabote Aspects of the port crue Calcobrad Unit Socorro Pro Median, 117 6 BAITLIY &RT Dne Docbron J : x 7 HOLLUD WC&CE Doxor 1034 Am J Phymol in 486 • TEAUTY W, U GOTT & K FOLDT 1953 Arch Pthe 203 9 MAMALL ) ) 1937 Circulation Rearch Ji 061 10 GOLLAR OG RUDOLPH &N & Oun 1957 Am J Phynol 107) 277 11 MATUR, GERA HALDO, REVICE OY AB HC00D & EB VADOVY 1956 Am J Phyu 1653) 515 12 CORDOBO&AH HION un 1955 Curculation Retarth 446) 575 13 SRL, G D G BARLOW I LAODIR BOUT Perodual con mama boa 14 V DC, $ 1957 I Metabobs Aspects of Troport AcTom Cal fembrane 106 Unry Wiconan Pre fiduco, 15 HECT H 1957 The Electropbymology of the Hart. Ana Ni And Sa OKO): 16 Buy JH 1957 Bnt lied Bolt 13 181 17 HOTTKAL, BTE E SOCILVO 1953 Am J Phyto 17h 312 18 JL &P House 1956 Circulation Rearch 1 332 19 ILALIM LI&OF HUTTER 1956 J Phynol 124.381 20 G Do Il 1957 J Phymol 1: 110 21 SOATDAN HJ 1053 Het Phymol a thermocol Acte 111 346 22 HAD, L & G Hrvey 1940 Acta Phywl Scand 11 347 23 Furo, LBA Gowali um 1912 J Bal Cha 14:35 24 ORL, F F 1950 Meter Pbywol a harmacol Acta . 52 25 Tiram, L D D D COTT & G J VOURO Cold Spring Harbor Sympo Quant Bal 1318 28 COWILDB LB Floonths Root 1919 Am J Phymal 123: 231 27 ALI, NOWS BALDT 1952 Am J Phymal 170 401 28 SITA, CRR OVAN IS &RCSANITE 1953 Cornie tro Romarch 1: 284 29 BANDW, G HPK London, J RR OVOXAN 1957 Circulation Read (4)1 419 30 LEI, IAGC NOTOTOOLON URR OMTREN 1958 Am. J Phyba 100) 64 ·EFFECT OF IOVSO THE ACTITI OF EZVIES DERIVED FROSI CARDIAC TISSUE John Jlants Department Buchni thari Cathos De Tuy stone two analytus of the various arton torolved in cardac contul mart ulo baro consuon the unique lettres of the highly uritable to use, ut mell u thone ut bolds in comerzon with other more anborra types of much tim Such a constantly nothing buck with no min as well as tunc neric Alppily operulary dependent opo iu in cartrondeat, etracellular ucl as intracellular Altantone is tbe vonir comportion of cruracellular Saad often have profund is This, as elevation or depression of it muriedly alter the electrocardiogram and the heart beat Stoke borncostatic control of the atroBalar fluxd composition kromplwhed ting many other Imma, lxing axl livet i particular the bypotbeus has been showed that mounty ardec Aboxarmale do not resa froch ardzac ne dy fancion prmanly bet, krandanty - a result of alteraixon on Winx compoarison of tttracelular food The wypergatic or on the other hand, entappatx fonctona of expucelalar ons opon the lastable Hauctors and membranes are documed by other contnbutors to the monograph Whether these pa tiert their madurace ty effrcting entypoc rectus 11 the sustace membrane or werely alter the check charge datribute of surface collana iz maties of peculation 19 anxellular orgina, seuls, and bacteriu ranom rom an etert profound effects at the 1 Cardmc care, like most other ones, control intracellular Lon compor con for the most part by actre trenger foechadura, hoch are membrane tomen in the fint utice There mochersun bere bera exteriency nye pia metve and we neon to toucheba bert made Thus, the vitu cuman of Ra from the cell interior eyed though the membrux x permeable to la Maxibed to the laterchesion mechancen located in Over the cost The Darpan Laganx phosphate, to wish the membrane dot roudily parvalde an act e process imarroga decal med mera berane trumport mechapp Evca the upute of stucor, bxcha ght be thought to let the suscle cell by pan cdiffon and then undsgs cautymata atuck on the cytoplasts, xaize to require a bugby Pecard, me myembant mechanit Is the cure of keletal muscle thro the proces cockered try any makers to be controlled by sun and order bormonet Onur kporlag of the trust your sections undertrag brine trasport DIATKROCTUS merger and a largely is the descriptore mere of der chopment loc mon coxents can be traced th ndoartive terzents, and the art of her bekk grumete u teraperature for rabainit of mbiutor coacratitu hu been dekbed The main difficulty thai manimulan cell membrana 115 f ·416 Annals New York Aademy of Sciences ant difficult to isolate and shen epart from the intracellular enzonment they have few mengirable enzymatk properlkes. ferhaps it is illusory to view the entire tranqori mehanism as occurring in the membrane the certy for Ibe proces u probably provided by the intacellular miymatk machinery and communlated to the remboide. The metabolk machinery of the heart is goured primanly to energy liberation By Rupplying ypproprately labeled Ribunto it has been shown that the main pathways of Delabolism are vu the Emden Bleyerbor scheme of shrolyot, fatty and degradation, and the atic and cele Hydrogen and electron trans port to orygen occur on organired eniyme chans within the mitochondria. All of the cytoplasma enzymes and subcellular particles are influenced by ons to a greater or Letter degree In a broad core cellular metabolum ama out 1 dou) (unction belo it la providing the cincy for the working muscle it is also maintaining the intracellular sonic environment, the high K+ Jlgt HPC- und the low Na+ und C24 In most mstances the only way to ime the effect of a ghien bon upon an enzymatk raction w to study the reaction kinetio with a pure an entryme as pomble thus eluminating ade effects that might also purious results тър - not of course the stuation in whk h the enryme a found within the cell There a number of compelmgo Rupponing reactions may alter the activity of Tevertheless, usa fint approach realus obtained on punified systems are at leur guides to the chenutenski of a reaction when it is into grated into a metabolic pattern From thus point of view ion effects on the major enzymatx reactions oncorned with energy release will be revened, and tben certain generalito all be documed Easymo Concerned with wcalysts Stort u know about son effects on the group than on any olber properly because so many of the enzymes have been solated in a crystalline or highly purified uute (TAJLE 1) Heart te besokinak u volated from bomogenata ube chutyme Tur 1 lox FJTORI ON OLrLine Fans Cene Atabx boute ATP Ig++ IONELL Qbor 6-P Co++ ++ ATP AS Fary metal Hay utal Hoook PbogpLoberou worden Phobotroctollow Ahola Thorphopbat debydragon PP tín Phoypadyak ortas Enokte Pyrark phopbatare Lacik debydrogen ATP 100 DP ATP 2 3 da PGA DOO ADP DP ZO Bitt Dare F- Cat Sr+ C++ NA U 11 NEL Zn CE Traty to per cent mbton t004 » Adenosine tripbocht (ATP) das diplombate (ADP), dpbopbopyidoro pickobide (DPV) 7 3 dphopbuh cek od (2.3 POA) Glocou 6 poophat (Gloc-6P) ·Bluntı Elect of Jons 117 tts in both a particulate and soluble form As with most of the himse it a activuta by light and the products of the reactions, ADP and glucosephaspute, art strongly way Both be glucose and the phosphate moieties of ghorphosphate art impaled in its intutátory effect Phor phofructolmuk also requran digt+ and the purtwlly punied brum curyme murally stimulated by YFI.+ slibourts, onliko crude prepunions, I * Dos &umulated by X+ kom It s got rahibuted by wolfhydryl Waching reagento wch a vodoucetamide reit a very ketve to Ahe mos It must be emple sited that there bevation portata to the bruin exttynse astuslar studos m enityme fronn bart have not been made Abolax rexmbles phobotrortokmake with respect 10 sahibstory areits beary metal bonis inactivate if hle vodoucetate don sot The murhatodd trok phosphate dehydrogettato continue real new Lacets of its structure It has recently been shown to coctam arx, uod the metal may be one Imk belmeen protein and the bound DP that reming with the enzyme depute repeated Torysalita son The other detaydrogoase a the glycolyti mchezrze lactic detrydrogener Dalmo e doc-containing truyurt, and it iken she requirm w other profi won for enrymax activity Enolak activation and Inhetatkan are reportant became they ilustrate acre way m wuch both cations and as ons nituence these phenomena Ignt the activator combisses with the wakubstar Food phosphate to forto magnanum duoropboquete Compten that the true whilator Although in and Zn++ in low oncentrators are better activators than ligt for the yeast curyme! mumed that ligt+ a the preferred activator within the cell, kance 1 more eflative at concentrations that are representative strellalar coxonen Pyrunt. phosphoferix abo di comandierable hotarne snportant because chamame of the farm chuyuses shown to require X* for manmur actrity Ia fact the slight reuernbeifty d the transphosphorylaxe moton could not be detsuurated at the startabatory role of Khud beea ducovered le now know of other reactions for wyntbainny phosphocpolpytuvait that serve to win the barre megy bestyr belke it and pyruvate hence K+ 0 longer plays eats a vital role in recrul of glycolym. EASTWS af am Cycle loo Seussettes for the group of enzymer have not been thoroughly in Tempted (TAJLE 7} 10 the unsyma are studied as part of the comptes mitochondru of sixh they are miegal perts, they are already in medium optimal for prezenta of mitochondre, geroally coolant, appropriate concentrations of Ulst+ 110+ K and phosphate xom Au they have been extracted and purified, mort informasion about the sequeretreats her pared Cryuubne oockenung Tanym, prepared by Odbor do frame rat beurt »u always andet mith K+ prezent In the way etaphoying phosphor trundtlax thu nu neenan ihn masytixe remuro K VIL far Alinity Yox dit aneettled the ion requirements for the modCoupy Keton In studying the same that is an ext of free ento ·420 Annals New York Academy of Sciences THI4 Ion Гch m Гута ти Surrot M Vary SopS or ETIOLOL Ceilte . tits Acuni TPV moonte" DIAL CODE Ouluttate decurbory luxe Pyrus onder Glatanic dehydrogen Clatex opurtu Indamine Oltacelame trup mer Socor) 1 Col thunder DIN DIT COLI branc DPN o TTY Pyridoul P ndoul T 70 is ilg++ Pyrdool phaphate (pyrido al P) thacta cod (upox) enzynne 15 order to mamtain optimal activity of the Krebs cycle it one creary to replenish vinou intermediates, particularly alıcetate The malk present in heart me and may be the Ley enzyme for this purpose It is polyfunctional the Ame enzyme atalyzes the formation of mallc acid from Pyruvato and C, and also the decarboxylation of oralacetate which ubmequently formed from mabaie by ibe all aty of multe dehydrogenase " Slangurore e required for both the COrfiring reaction and the oulacetate dearboxylase uctivity Inhib tron of the decarboxylux activity by malete would seem to be a ureful protectivo device Krving to maintun the oxalacetate for the vual condemnation reaction with acetyl COA The omple pyruvic Oxdure reaction that the main link between gtvolyes and the Krebs yule involver one or more enurymes and four cofa ton A preparation from pig heart rus tested by Menang dinte formation in the prescate of added coodanung eryme Jig++ and Krone ben teked for its bility to form acetylphophate in the prerence of added bacterial Irons acetylase Jig++ us not required Trom thu it may be concluded ibat the oudax docs not requirt Vigt+ The requirement for other com mu not adequately cumped Hart tue contulos considerable mounts of glutamk dehydrogerus in enzymoc that might functon to furnah a-ketoglutante hence another Ley substrate that serves to maintain the Krebs cythe Sullee daha e thonn that liver glatonic debydrocene na nnc metalloentyme hence it a powable that the muscle cryme also contains ADC. No other son requirements be e been reported other than \Ht xhxch 1 Rubente in the ration The innovams have not been rufficently punted to permit a potous analysis for tnce metas hon ever It has recently been reported that ligt Almulata the activity of glutemkoalaceti trimeroase us much 4* 80 per cent above the acti aty obeyed mitt abectie Role of lors 1x lrytalic Reactions A laking of jo repairements and other moduying effects of ions in entymath reactions serves the useful purpose of emphasring those that must be present In order to cure adequate activity In addition to consderations of speci- ·Jlantı Efied of lons 421 futy it would be desirable to delimit the coox mtntrota od korus required for producing an optimal effect rusthesore it would be very darable to knon har egenvon works in turfreution or BT, etxeths rould be useful in mainions on the on rolling mechanwa m enrymt rections Definitive ecplanation of too effects on the tres are len w, at ibe mask of reiteration, move reprexataine tbeonas all done be briefly cummed It is awful to clasſy bonus commonly moved in thisymatic reactiona mto the follow my categond (1) wes that are integral parts of an enzyme complet axentally and secrubk, and that itmain with the enn de brough externe ponfatra procedures (2) Kas that are bound to the protein moviy olan ensare and often to an anonied colector u wel tot ere more or leu dowcx ssble and cookerWestly must be present in tbe free state mode definite conception (they toay also be bound to cofactor of mbante to the first Innoce and ther, madaniy to protei us a metal soberate compter) and (3) kons that are bound feebly us at all, and krve mainly to prondelor uwe ervondent for the tnsyme (1) lex as Iwiera) Esrsyne Components Thiene coraplet crurymat batu afien been called the true metalloqxolers Typral reprekbutives coolain rod 15 in the beme proiets and ra certain flavoproteiny in the former und unclergoa reserable onderson reduction however it uncertam bether the iron undergoes a malene barige la re act and morng the flamprotan Eamples of enstymer colamus, copper are tyrome and abo we recently doomcred fleroprotem. Carboox nhydrake u be denk -contamnog mome more rreth castory popular and a number of debydrogeoak hart been curacterised os anc metaloprotam* Jolybderrum-containing tarym Rhos anihide dudare, mod artrate redoctus bolal ma tha category Except for certain bcnc pries, par kon ledge of the mature of the bandung between metal and prorano manger Studear on yer alcohol dehydrogerum and much lacuc nad dehydrogezzax bare naked the fact that there is an mtos RIX fery molecule of bound DPX which suggests that the metal comreme wnb ibe binding of cofectos to protein Perbapo one of the bat crushes of the ones which encui sought act usu bodig art and at the more une participate n the analytac function of the earyme come from modd Hua Hb pyndaal, the cofactor of the transaks Baddskeyo has shown that solutions Commune opindotal, pyrdoumine an anino und a ket xid, and detal katus Cost form med coordination ompounds of the type united w nourri In the imido and depola by R tum rind wib the pyrkosal pest of the campki and the Leto aad Rith the pyndoumide part The ben onderzoen election and proton roarnngainenia ** bona bereun Rus become the leto kod ad R the per to sad Podukty that the meul could be coordinated t * curyme that mithi represent the intercorersion that occur in an entynsulkuly atalad trinarumattor Vafortunately the in Don abundant inmamma h Dot been prepared as #sle of efficant party to permit rem ble avea ·420 Annals New York Academy of Sciences TL 4 LITICE Oy Land TAT SOPPORT THE SAM SCRD Or LX METABOLMY TL Caus bent Jualdory TPV JIH Blac Dulomate alc cautyto Outut decarboxyleso Tyruik odun Chatum.c dehydrogenas Glatam put them Glatac-tlamo tren MK Succinyl CoA tudom DPA DPT CW bporc DPN OF TTV Pyndoul P Bandoul P 20 tech fyndoul phosphat (py ndoxal P) thotk wod (bipom) order to mantan optimala tvity of the Krebs ycle it a necemr, to replenish various intermediata, particularly lacetale The Toalic enryme present in heart tusse and may be the key enzyme for the purpose. It polyfunctional the same enzyme atalyzes the formation of malic acid from pyrunate and C1), and also the decarboxylation of oublette »bach b Aubweruently formed from malate by the activity of mali dehydrogener SlangaDene u required for both the (Orfixing reution and the onlacetatc decarboxylake u trvity Inhibution of the deurboxylase activity by malate would seer to be a reful protectne devue krving to mountain tbe oralacetate for tbe viul condensation reaction with acetyt COA The complex pyruvic axxase reaction that the main link between pyrolyn and the Krebs cyclo involves one or more ensymes and four colact n. A preparation from my beart su tested by merrnng atat formation in the presen«c of addede odenang cnryme, light and K+ On When toted for its ability to form acetylpboopbate in the presence of added bacteral transucetylaxe Bigt wus not required From thu it may be oncluded that the ondase does not require Sigh The requirement for other tons was dot adequately cumped Heart torso contains conoderable amounts of glutamic dehydrogenas an entyme that might function to furnah a-ketoglutante bence mother key subuinte that serves to mmtun the Kraty cycle Vallee d al have shown that liver glutam dehydrogenase B a un metalloenzyme hence it b powable that the muxle ensyme il contados anc. No other ron requirements bave been reported other than VH+ which B i mbante in the reaction The inmummies have not been fufficiently purified to perrait e ngorous umlys for nuce metals home it has recently been reported that Vgt stimulate the activity of gluten.c-owlacetic tnmuninuke u much u 80 per cont above the activity observed in its beence Rod J Toms 1x Pusymalic Rodions Alating of ion requirements ud other moluying efects of lors in crusyonatko Tactions serves the useful purpose of emphedring those that must be pretent In order to assure aderate actelty In addition to comodations of pea ·Munts Efect of lons 421 fiaty t would be deanble to delimit the concentrations of ons required for producing an optimal effect Furthermore, it would be very darable to know howa geen on works in a specific reaction or Hon, usce this would be useful un investigations on the controlling mechanians in enzymatic ratom. Definitivt aplanations of an effects in thu cu ut few mod at the nuk of rateration, som representative theone will now be brely on mined It s useful to clasify lona commonly mvolved in enzymatic reactions into the following ategoria (1) DS that are intenl parts of an myme complex, encatully Dondoocuble, and that reman with the entyme through etmeye porfiation procedures (2) mons that are bound to the protem monety of an altymne und often to u usocuted cofactor u well, but are more or les domocable and consequently must be present in the free state in come definite concentration (they may also be bound to cofactor or mbunate in the first inance and then secondarily to protem us metal-rubstrate compler) and (3) sons that are bound feebly dat all, and serve munly to pronde a favor ble environment for the enzyme (1) Jous as Iwegral Exsyme Components These complica enryma have often been called the true metalloproteins Typical represent tives contam iron, u m the beme protens uod in certain favoprotein In the former Top undergoes reverable oxidation reduction however it uncertan whether the troo undergoes valenco change in re actool involving the flavoprotens Examples of maymay containing copper dre tyrannar rod abo some recently decovered flavoproteios. Carbon unhydrex the damic anc-containing eryme more recently arbory pepuder and a number of debrydrogene bave been chinctered us on metalloprotema " Diolybderrum-contaming enzyma sich u mnthme andar, and nitrate reluctas also fall in thu category Except for certain heste proteine, our knowledge of the nature of the bundmg between metal and protein is mager Studies on yeast alcohol dehydrogenas ed uncle lactx iad dehydrogenaxe have revealed the fact that there is an atom of ac for every molecule of bound DPN which suggests that the metal concerned with the binding of cofactor to protan Perbaps one of the best enamples of the manner m which metal might act ur bonding 4gent and at the me time participate in the catalytic function of the coryme comes from model studies with pyrdoal, the cofactor of the trusammuret Beddikey has hown that solutions conting of pyndoul pondomine en amino acid, ket lod and a metal och u Cutt form mixed coordination compounds of the type illustrated in TIGURE 1 In 4, the umino acid denoted by Rhu rected with the pyndonl part of the comples and the keto uod R with the pyridonmne part. This then ondergoa electron and proton it u shown in wherein R bas become the Leto add and R add Postulaung that the metal could be coordinated to might represent the nterconverson that accun in an enry inapambon Unfortunately the is mort abundant to Dot been prepared mu state of sufficient purity to permit 10 ·422 Annals Nex York Academy of Sciences "HOOC-CR DH ch CHE HCN RCH 01 COOH COOH сн. HOR CH b RCCOOH Footu 1 metals however a punfied tytoane-ketoglutante trimmines from In er has been found t contain copper # With the recent improvement in glutamko apartxctrinare punfauon," it may be possible to enamine the problem of metal requirement more abally One may speculate that the cliroox depletion of a metal required for the continual resynthesis of a metalloenzyme could lead to procreative duronu t100 of active enzyme and that ultimately it might become the limiting step in energy relair Indirect evidence that the an occur in a specific enzyme Fyrcm has been presented by aller a whok studies on patients with pontakcobolxc arthom led them to cooctude that a cooditioned ainc deficiency develops as a result of the recurrent bouts of alcoholen that land to progresave dımloution of liver alcohol dehydrogenaxe actity and, altumately of proten metabolik (2) lors del lloc o Cwpane 1dion The lona moat frequently encountered us thaton m enzyoutic reactions art 18++ Dott and ocagonally Catt A common property of all these their ability to form chelato frutura with a venety of subanta, particularly thom containing pborphate or condensed pbosphate group. Thus, Dight lo in almost univeWi component of ractions involving ATT « AOP Bian gadete may also fanctloo In tbex reactions, frequently Dot us well as light On the other hand, Joo much more effracious in the decarboxylations of A-kato ods There lors payı dual role in cheating with the state they star the electronic #ncture to us to make certain groups of the Abante more vulperable to atted and at the nume time, they furub Ink to the protein in forming the easyme-tubetrate complex ·Nunta Elect of Tons 423 0-C-G-C-C002 0%{->CHẠC, Proten 0-C-C-CHE Protant TIC 2 A cu instructive entrophe #yun model system, the spelaialdad decarboxytation of dimethylosaleceuse cod mehnatod by Stenberger Land Wexthemset " If one projects theu romult to the entymentalymed che carboxylation of oralareix not the mechanm shown LA Foot / ould be opernity. The metal ma tha se mannerBy cheating with the acid through the carbazyl and educent kelo group insieme zums un electron whift from the carboxyl (urbatu, thus water the CCOO bood and i beating CO, 'The resulting enols then cleaved from abs compla by a proton, and DYTUVC Acd a fomed. By brading to the protein, de mange chelate unbergom the inciso very rapedly in the honenymate dhe tymon, pyndinc promoted the deaastotylist, thus gring add110k wupport to the proposed moxchange with coxyme in the use of wanne 18+ houred phosphorytoons notably those ac CUINDE in the fructokinar cod poptofructokursuse macione, Lardy mod l'art have poulet out that the ATPAL nato s very portant If this Auto w ceaed to the greater than i proprestrely me odation ** obectved, promurably because the uxcomplesed ATP inafered to the beading of AIR ATP D enzyme and thus denuntthed the fate of reactie The on technoeg of the effects not enom n bat gasa i wooki fpeer that be bending of Might to the prophetate group members the termal staan phante raore racire sod like to promote the banding of E ATI' 10 eurymt. Klatra as desconel ruout type of gooien binding with picta completer (3) lo awr Fonlide ik (* * of Subaret ( maler nuk fayne The mandralent 2, K+ and NII me the mout common emplo of There are les routes in het bos wors are absolutely re you! Only thoa cattomed by potank w, poubiy ·424 Aonals New lork Academy of Sacaces condensing enzyme among those that we have been considering here Seren! othen are stimulated to virou ertents when these lors ure present The manner in whxh they exert their effect Beczally puzzling once they do not keem to be bound to any importedt atent ather to cryme or rubente. Honcver Carr employmg an electrometnc method hus udduced evidence that it bound to i mall atent sith certain proteins Although Mat also bound, the extent of binding us wally smaller than with K+ The difference between K+ iod VH+ on the one band and Nat on the other bave been emphasized over and over agun Quite getenlly when K+ stimulates i action, the addition of Nat to the system wil inbiblt it Het thu phenomenon is not a typical on antagoniem Thus, Kachour and Boyer have shown that Na ul stimulate the pyruvic phosphofenza reaction slightly even though it everts a por crful inhibition on the stuulated reaction From their kinetic data theke uutbon coclude the boding of K+ to substante und muggest that it in some sort of combination with coryme, while 3g++ the other indepenuable con m the reaction bound initially to substrate (pytophophate morty of ADP) The me preference for K+ over Mat nu well shown by Gamble in bo work with lover mitocboodria Carefully prepared mitochondra with their osdauve phophorylating capacity mtuct will biod 25 tunes more K+ than Net per mg N The K+ bunding apaty is still returned although to 1 bemer extent in mitochondrial fragments prepared by digitonin treatment, which also preserves oudutive phoqphorylation However mecbadxal frumenta tion, whkh destroy the latter also abolishes the K+ bloding ability It u difficult to ripuuze anything other than dectrovalent bond farma tron between the monovalent alkali atons und proteins It u known that polyacrylate migomolecules in solutxon an uncute conoderable amounta of alkalı metal on Extereon of the finding proteins should also account for the high selectivity for K+ or other utora In FIGURE 3 u depicted a model to account for K+ binding and the way in which the fofluencorection veluty in an enIVIDUTX system Jlogt of the enzymes we have been decuming bear a net negative charge at intracellular pf Thu cburgo would tend to repel a highly negatively charged Ribeinte ach us one with a pyrophoghato group. Completing of this group with Vg++ would reduce its electronega thvity but not abolish The net negative charge on the protei to contoh uted by many arboryl groups and, ln mme au by phosphate group us nell. If these accur within the proton structure (within the bela coils or clewhere in the martor) ation would have to pereinic to the ate, od pendetan auch u hydrated se iod mobility of the catron would influence the penetration Apocution with these interior wies would reduce the net negathe charge on the protein and permut more facile approach of the ab ante compler to the entymatically acthente The might or muht not be the Ad u the tincting nte. The selecthaty for ilions of certaid doncmxona brings to mind a cus of compounds alled Lathnte formed by loching a guer" compound within pues formed by the union of Kent host compound. There * con den ble selectivity guests if they are too lurge they cannot be enclosed ·425 Muntz Effect of Ions Net negative charge Less negative chargo Breton ecotel k* on the R-O-P-O-P-09 0 ---- For 3 und, if too small, they an slip out of the age like structure Zeolita, bentonites, and synthetic resins ha e been comidered to be mold macromolecular structures of the type it would not be umut extend the analogy to protam in solution In concluson it should be emphared pum that even if we accurately define the won requirements fo to polated mayme, the alumet goul mot be to understand the factors that regulate to activity in IMO There 1 imple evidence that many enzymes annot be worlang at manmum reloaty thin the cells Consequently even if the KO environment » Dot optimum there may still be sufficient reserve enzyme activity t meet the ordinary needs. However odo consequence of a chronk hon imbalance may be programu donnution of thu rexive to pont wbere adequute function mpaired Rderace 1 ROUTED A 1954 Activa tnport und vertoo Stop Soc Fopul Broil 108. 2 CHIL RELA Sou 1950 y Bal Chee 104. 273 3 MUI, JA 1953 Arch Binche Biophy 135 4 YALL, BL, LH00. J ARTEN & WE TATO 1956 J Am Chen Soc Tor 3675 WAWD O W non 1941 Bancheta Z 110 31 6 BOLD,PDHA LADY PH Pan 1942 J Bal Chon 14 673 1 Oczon, S J R S xtu C SV 1931 J Biol Cher 1: 691 8 Yox Kaun RH 1953 J The Chan mas DOCXXS R A CLODIR 1990 Arch Bachcom *079 EBJ JO Som KDY I 1956 And Bocbero Bopbyu 11 Vorro, La 1956 J BW Cha M H7 13 ore 1956 Babe J : 352 13 PLAUT, E&S C S20 1951 J Bal Chro 307 308 14 ET, L LF Nyam 1957 Boche et Boobs Actu *- 40 15 Sro, TP EB KUD & VINY 1956 Farymas Units of B Suroitu ipd Function 417 sadar Pra Na York, NY 9 10

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16 ROX S11&X KLOTZ 1057 Irch Bocbem Biopty. 67 161
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21 KOGU 8 AD (AMMEN US O 1952 Bol Chem IH 541
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24 VALLE, D L 1935 Wasco Irotan Cheon 10- 317
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·INTERACTIONS OR GLYCOLYSIS AND OXIDATIVE PATHWAYS E Racer and S Gattt Drum Reno Pelo Tu Axl Roroch laders who Cxy No Ya, I* Ya Tuh, Y Y When I wu approached to purtkipate in this sympodom a metabohc jac ton in beart contractility with a review on glycoly and the hunt pathway I attempted to convince the program committee that my grannce of heart connaiſty 1 Umost equalled by my ignoniace of beurt metaboban Тъв did not chminate me un contributor to thl monograph since I cannot pak about the curbohydrate metabolom of the hart, and once tho ba sympopum on the heart, I will diecus what I bopo can be alled the heart of arbohydrate metabolum When we attempt to rudy the role of metabolle factoo m the phypology of en ru ruch us the hart we try t lam as mach us we an about the enzyme yeters that function in that organ In order to do mo, it becamry to dwupt the organ structure and to bolate the individual enzymes Enzymology . relatively young aceDCE To panphrax Bemurd Shaw "Youth Be wonderful thing it seems a pity to wuste it on the purification of enryma However wolation of the enzyme rystems has been the most nabonal approach to the study o mechanions and now that easymolog begins to approach matanty we find that some of the en-symologists are willing to part with some of the pun. tangul view of thefr scientifi adolescence Previously enzymologati thed intact te cells merely us starting matena for the punication of enzyna The more brutally they could treat the topic, the better they liked it. With the recognition of intracellular Aructures auch u mitochondru and microtome u metabolke organ, the crymologista developed a more gentle upproucht th cell Not only did th> open I DET world of multienzyme rystemet them, but they ould now take advantage of the structunl dutribution of enxymes for purposes of purification Among these enzymologats a few have tam a further step uod tuned to the study of eprymo intact cell The difficulties that confront them bare ure quite different from tbove encountered with purified entyma or even with ell-free multienzyme rystems In order to mahate observations that they male with intact cells, they must know th me yor alt mative pathny they must learn what limits the rate of ach of these multipryme sytten they must be aware of interaction between the majo rytems and of the DUIDTOWN ude ractions that erat in the complex metabolle machinery of the cell Per haps the generalining will become clearer hen dal with a more concrete problem the tilization of glucose Tbeort duanbod tu paperported part by Great C 363 fra thN troon Cance Istitut Pula Halth SerraBelke lid and in part by prot treo the Juze Coca Cmil llenalled for Mechal Road, Na Ham Cann # Fior of the Juc Cortio Chile lorul rund for lletal Rosarch Mogata di Depurtment of Bootbountry Hudh Herr Urranty School of Jefone, Jeunkt, Ind ·enzyme level 428 Annals New York Academy of Sciences There are three major pathways un inmal tiguas responsible for the break donn of glucose to CO and water (1) the glycolytic pathway (2) the triar borylac acid cycle (TCA cycle) and (3) the oddative pentose pbospbate cycle(P Pcycle) Thxatabolic systems are Interlinked with the metabolism of fats, proteins, und aucker ude in an almost continuous Dei work of eniye catalyzed reactions Thoxe relation ul not be discussed here. Imstad, I shall concentnte on phenomena of intenctions between the three major multienzyme syzicos af arbobydrate metabolam and point out some of thels ardilantio and differenca Tint I shall discuss the interplay between glycolyus and the TLA cycle The enzymes of glycolyus and of the TCA cycle are structurally spanted inade the cell und me an ume that there is Utile direct mtericton at the Horever both systemas require nucleotide cofactors and loor ganic phosphate A phenomenon much us the l'auteur effect iodantes that Intracellular metabolism involves competitive proces In our laboratory Wu has studed the phenomenon in recent yean in intut ancien tumor cells, and we have abo investigated it in a reconstructed system consisting of liver or heart mitoctroodra and . muture of punfed glycolytic enzyme In the Kudies with the recoctructed gutem it us porable to vary at D1 tho indrudul omponents, and I shall give a fen cumples of the competitivo phenomena obuerved in rocb esperiments It s now well establohed that carefully prepared mitochondria require ADP and loorganu phoqphate for respunton To permit the study of competitive phenomena, adenine nucleotides were used in aulythe amounts together with un ATP regenerating system The effect of varying ATT concentrations in the presence of creas inorganic pboxphate berokmax and glucocos shown in TTCOIL 1 In most subacquent cxperimente, 3 to 5 X 10 - JI ATP were ted which just belom optimal Instad of glum.ome and herokinase ATP.X was added to some crperiments, u shown in GORE 2 We can see that a propounced stimulatio of respiration mus produced by the addition of this entymne A romperson o rotus 1 und noULE 2 tbows that, wbile there was hardly sny respinton without added adenine nucleotide even in the presence of glucose und hexobimate corsiderable respinto nas obtained when atalytic amounts of ATP rere added alonc ritbout an acceptor system In fact FLOUZE 2 y not entirely reprezentative in this respect as, in most copertinents, only two to threefold stimulatim tu achieved by the additton of berokiare or Apyrex le mixpected that respiration in the presence of catalytic amounts of ATI might be due to mitochondral ATPig activity Such an activity sus readily demondtnted in our mitochondrial preparations I should say 'Tadily" beai other invertyalon hair ported dlhouliks with #clutade met purements Albmursued throogbout the yer !7 Pood App for makromet souphal and une dystrybut DI Irlor obud bobupno tockowe ad in phophora almockulle 3p for glycey Webs de proport 1+p for my thror + phosphate Oopfue din opphete ! 6 p for frextose 6 phreybu Spfor wedohet boer Johannes, 16 loro posocomie Mr for fronton 16 iphoptat SDP for dobetubboe 17d pbophet Plr for poporod gate EDTA' for by lent un tetrixtut PPgd for patos pabai sck TCA pek for Inarton bc sod gck.

·Racer and Gatt Glycolymu and Oxidative Pathways COXYGER UPTAKE (MATOWS7 068 05 10 *W ATPAN Fon> Lord of ATP Concionuort a respingat natochoty Excllasbary Aktuetan the one of20 00 odebutina 20 wool of platom? 50 modes of phompot 10 note KOI 50 arter of tancome, 10 lot of Sr 30 onts of betakrat 43 o rat inet n cochondra me AR 14 The TE Watang repromater for 60 ton DC of mitochondrial ATPase and wc too bure countered them. The therefore de cloped . Beway procedure Lar ATP that not only has the admata that it an be arried out at mort phymologiaalkeel ATP but that it n more produabic The stay contabus an ATP regenerating Tem conting of phoghornal pytovate magnesium, an cucen of pronte mare, ad ata lytic acontrol ATP Inorgasm phophate formatie was found to be propor tonal to the amount of moochoodru o apynx added Furthermore miochondrial ATPax samty maaared by the prxdure u found to be mama umes better bad when me gured by the coventional method without ATP regenerato, us abown in ta I The most plek moon for the dom arpancy in the tro anuyo mu that ATT** * mhibited by ADP In this nurt unstance an obvious explanatxa turned out to be a corect sose. ADP a COKYOltrona beyond 103 u found t tabibit mitochondrul ATPK An obecreation made previocaly by Kxley and Kielky The inhibition of 4TPU by ADP my well reprekat s regulatory rectus for the arrior And apparrthy agdul ATPuk acry of motorbroodru. Just us tbe addition of phosphate saceptor antens manco ropion or mitochondria, the addition de competing phophorylating system, sit us ATP-omine tubonphany kve (Fourt 3) or ghrolni (T! 2) was found to inhibit reppinutki The inhibitioa no revered by add ton of ecen ADP or 2 himropberal, hech esdicated the operatia de compenne mechi ·enzyme level 428 Andals New Jork Academy of Sciences There are three major pathways in animal troues responsible for the breal doma of glucose to CO, and mater (1) the glycolytke pathway (2) the triar boxylx add cycle (TCA cycle) and (3) the oxidative pentom phospbate cyclo(P Pcycle) Ther atabolic syntes are interlinked with the metabolam of fats, proteins, and nucleic acids in an ilmust continuous betwork of enzyme catalyzed reactions These relation will not be discuued bert Instand, I shall concentrate on phenomena of interactions between the three major multi enzyme systems of carbohydnte metabolbum and port out some of their milantic ipd differenca First I sball discuss the interplay between styrolysis and the TCA gde The enzymes of glycolymas und of the TCA cycle are structurally separated oude the cell and we can asume that there x little direct interaction at the However both systems require nucleotide colacton bod inor ganic phosphate. A phenomenon puch us the Puteur effect indicates that intracellular metabolisma mvalves competitne procteurs In our laboratory Hlu bau studied this phenomenon in recent yean In batact excites tumor celb, and we havo ulao investigated it in a reconstructed system condimg of liver or heart mitocbondra and a muture of punfied glycolytk mnry In the Hudies with the reconstructed system it was poruble to nary at all the fodividual components, and I shall give a few enmples of the competitive phenomena observed in a ch cyperiments It now well established that curefully prepared mitochondria requre ADP and porganic phoxplate for reginton To permit the study of competitive phenomena, adenine nucleotides were cred in atalyta umounts together with an ATP-regebenting system The effect of varying ATP concentration in the presence of excess morganic phoxpbate herokines and glucose born in FIGURE 1 In most subsequent cyperiments, 3 to 5 X 10- JI ATP CTC used, which o fust belon optonal Intad of glucose and he tolumax, ATPase us added 10 sonde experiments, us rbown in noin 2 We an see that a pronounced simulation of respiration sus produced by the addition of the enzyme A comparison of TOUR 1 And IGUIE 2 shons that, while there us hardly any respiration without added adenine nucleotide, even in the presence of glucose und herokinare cannderable respiration sus obtawed ben atalytic amounts of ATP were added alone without an acceptor system In fact TIQURE 2 pot entirely representative fo the respect 4, in most aperiments, only two to threefold Almulation us achieved by the addition of berolinax or apyrusc Tlc aspected that respiration in the pretence of atılytx imount of ATT might be due to mito bondral ATPe acti ity Such an activity or readily demonstrated to our mitochondnal preparations I should may readily a because of bet invatal have reported diffx ultres Ith accurate merrements Aber YJ hrobootika 17 Pund IDP la denoroc trlponphat and ne lobosphat Di II for oglurd dropbo rube achoutke od 1 porqam Ruck for th cetaklets de 3 people rup for ory throne phrabi Gop for boyha 1 6 p for frect * 6 phoque 57 p for Kubbop ko 7 pbopha OIK I loro phosphatocoma! IDN fæ frocton 1,6 dpat sDr for adobepilom 1 7 dipbophet Plr lo phospod prsa EDTA la cthylord amine tetructut Prake for peator pbohat acte ICA Grete for incubory pod de.

·Racker and Gatt Glycolysis and Ondative Pathways 431 OUTM andan GSAXO so 100 200 100 J00 HG ATP-CREATINE TRANSPHOSPHORYLASE TOOL 3 Inhibition d Atochondre pintloa by the phopbocroute cremelor metom Each Wurtary tank controed in Goal volume din 25 pola Iphoghat 10 Mol al bondme 10 soles of naturmate 20 mole of fructo 20 mmolen drá,03 my of DPN 5 mol of MCL 033 pks ATP 10 mg opboghormati 0 6 Onits of boron 48 m al rathri mtochondre opended mol of 025 1 mental contamy 1 mg/ml patru EDTA and ATP autune triphosphoryl (creatme) umdicated LbOTE The Gol HB74 The voics on buen for 30 DLD At 27°C TU2 LATION OF ROTATION GLT TU (CRADIO ETC) Adittene Corytale rak 31 SO Horom Herold + yolytec 2 11 14 11 T ful role of 2 ml cerdded 20 moto al tratongte, Dolu of Kpbron 60 polo of hodine 10 mola ata 50 Dola of pacote 10 dolu af Lety dropone 12 tout of herokee Id 1 2 mit fpompboirecto In the open mint 2 d Toda fnction from about made me the part of your moo Thor rt shikon for 30 minut 30°C It mu tlo ponable to imitate the Putear effect in these reconstructed multienzyme yetems A shown in TAC 3 the formation of lactucd wus mit edly inhibited wben reparing mitochondita eft added t głyrolyzing system The empalment u curned out with limiting umounts of dende nucleotides and creas of inorganic phosphat In order to paform similar urperimenta in the presencm f limiting 10ount of Inorganic phosphate tuu kecemary da me sytem of glyrolys that proceeds under exch cooditors tho in TL + glycolym an be maintured t low Me concentrations by cl th correct amount of ATPixe, Vlthout 41Pure Little lectx uod ·432 Annals New York Academy of Sciences TAIL 3 PAT EMOT DE ARMOID SYID Op quake Leam Glycol IX, Xam, no matochoodie Glycoh the tem with mitochondre 9 5 6 39 Ivmir o serpenmaats gerbo andation quotient - 3] Each Warburt va contined in tral volume of 1 ml pole al htklo 10 moles of Kalotest 23 mol a phophut 20 roken of KCI 20 Doto al nuctor 03 DE O DPN 03 mole of ATT Smokey or C 0 25 units of becomes (emused th phucove u robent ), 06 to poor belacionar 32 unts of a kolex, 18 mits of cyceruldebry de 3 pboopbat debydrogenas 123 ct of pbop boglycerut ibu 21 mits or phoxpbody cetat mutes, but almolux 38 mil dl piruvate Line & 7 umb of clic debiydrogenix ind o 5 anodes of 23 hpboopbody cerut Tbc reach hoch us incubated the peace of mtochondre, contained 71 mg machandral proton mapended on 01 ml of 0 23 31 manto contamint 1 mg/ml of Deatra) EDTA The final PH u 74 The cocks et cubated for 30 on u 17C und de proteired by the addition of 1 to al 10 per cent TCA 610- STUE MOLED A 02 0 ठळ 01 OS APYRASE UNTS For 4 Efect of pine an scolas Fach test tube coolaned m Anel volume of OS o 15 prole of Dogdun 025 poker a formante 025DPY, 12 moks of broctox 10 Dola al Ka 015 polo of ATP 25 Doles of MC, 03 or molad phophat 0 17 mts of backunge la mayed the place or robert) 03 ants of phon porruetokone, 24unts of adoles, 24 mits douldebyde Jpboplat dehydrogenas 4 wts opbop boptt kmax ut opboplodytett molne 2 st or coax, 125 mud pyruvu ne 4 urt of lecte des drogtour of my china 0025 mola al 2, diplophop cerut mod apre in Teryamotnus o thon in the fourt The final ph vu 14 The muture An incubated for 30 m t IT C and apro tand Wibe edition on a 10 por cent ITI os cypeted in view of the need for sporgadlc phoqhale for glycolytus while ce of ATPase inhlher glycolyu because it effectively compiled with hegoak for ATP Because of the nther narros noge of ATP're activity optimal for glycolgada, the experiments on the Puleur effect in the reconstructed syutem with lusiting ·Racker and Gatt Glycolysis and Oxidative Pathways 433 LR WOLs) DO como a NOTION om WITHOUT WITOCHONORA som OUT WITOCHONOR HOTH WITOCHONOR) TOCHONORM ÇETE TOO MITOCONOMIA WATH LITOCOKOM O 10 10 10 40 0 O 10 50 MINUTES T.Cu 5 Kinetn ltboth colytic man ut low denne ochooddas ind prophet in the price ad beend of motorboodra T buks contamed ach in final rolure of 53 ml 165 mola al DL hoone 110 mmole petamet 275 mg o DPN, 165 mol ATP 110 moto of KC, & mole di poco bol al pompbat 2/3 pokes of MC 26 ta beraktis 74 alto a phopbrugle.com nomen 33 umts of phopbo froetako 30 minuts al wklokere 26 Out y do 3 phopbat delantero 52 toplumpbactent lines 23 cm of people com sout, 21 mit allem hectic darydropol 001 unter0275m? I borobogtyn ud 055 ml 220 25 11 punto my mutochondra proto ruspended in the mo mofium The foal pH 14 The testo oro balm ut tand the buna poobed was the same 21 of the action murat petied unto (1) 1 ml of 10 po ccat TCA eod ()1 03 Be(OH), teen 1 od bol zoso, u cihad The muloos are reported for 1 ml oli incubation motore and are corrected for evaporation Badeby IDOTONK phosphat concentrations were more difficult to execute, uod it mus Decemory to follow glycoływ at vanou tim mtervale us shown in TOURIS It can be seen from the ngot-hund portion of TURI S how th presence of mitochondru nhubius both lactate formation und glucose upute from the left hand portio of the TIGR 500c cun ded ce the rouson for the inhibition In the presenc f mutaboodne, inorganic phosphate » depleted o npdly that It * not avulable for ather glyrolyik or ddative regeneration of ATI It must be understood that we look upon these ctperiments with the recon structed system oly u models for the study of intenction of multisyne Sitem Althoogb se bave produced . Crabtree or Puteur ddect with thero enrymes, me cunnot tate that these pariments are represcotative of intracellular evento once ned not know what limiting in the intact cell und Ance competitive effects can be obtained in the cell-free sutom by ether limit iny Pi or ADP or both Hornyet the bort cipementi mablad s t upder Hund under what conditions competithre phenomene take place and prepared for th peit stop the lovestigation of the intact celle Before turning to thu upect hom ever I wall decu some of the mention between the glycolytx pathway and the Peatoc pbocphat cycle ·431 Andals New York Academy of Sciences emate as 200 00. (a) Fou 6 Serpenice daubon modation of phacone dari TCA de The glucose molecule whxch Inreb via the Emden-deyabol pathway pht into 2 molecules of pyruvate In macht manner that arbotu 3 and 4 of glucose become the arboxyl group of pyturite During the subsequent och datlon vle the Krebs cycle the arboryl group of pyruvate i detached bot, then arbon 2 u ordized, and finally the methyl carbon of pyrunite bordized With reference to tbe poucose molecule tbe kequence of oudation arbons 3 and 4 fint, qurbons 1 and 5 secood und arbon 1 and 6 last (TOURC 6) In the course of the persone phosphate cycle shown in ton 7 urbon 1 boudbeed fiat nbubone-S-phosphate, the product of oodatuon cycles back to glucose-6-phosphate Carbon 2 of the orginal glucose now carbon 1 of the penly formed glucose-6-phosphate and again is oudised to mbulone-5-pbox phate. After Edother recycling of the pentose phosphate, carbon 3 b ondeed in the ume manner after becoming arbon 1 of the benly formed glucose-opbospbate What remains of the orginal glucose is a molecule of glycerulde hyde 3-pbosphate which converted to duhydroxyacetone pbosphate and condensed with another troue phosphate to FDP FDP clared to fructose- 6-phosphate which re-enters the cycle and thus arboos 4 5 and 6 are burned by the same mechaniom. Ilc and se, therefore that the sequence of arton audaton diffen, depend ing on which of the two altermative pathways of glucore uddation functions. If we attempt to evaluate the relative contributions of these pathways by malog CF or Crlabeled glucose me must realize that we are dalloy with nate differ ence in comples multuensyme systems In cells with very active translctolase iod innalolax there an occur axch e apd Krimbling of the arbons of glucose that it becomes difficult to evaluate pathways by the use of glo.com labeled in ungle carton. Fortunately in mort mammalso true the PP cycle ba slow enough to permut qualitative Hudies of the mode od glucose utilo tion by intact cells but in view of the cochante reaction awyzed by tanke tolx and transulolex it seems alto-ort Impossible to urive ut ucunte quantitative conclurons Let us consider the kind of intenctions between glycolyabs and the P P cycle ·Racker and Gatt Glycolysis and Oridative Pathways 435 COURA Iruar 1 he Pepe 2+-+-POP3 F * IMA FDE Inc Pro w Show . Recht-02 The condamna pentou pboophut oda. Four 7 12000 Polyroonide O nomb-pbogtata 0-6-2 P-6-9 T-b-D Pentor Mbogate TOP LE Pune IT 20 dolu (GDP) PP Gole Iotomediate (0-P L 6-7 P) و ململا 0--P Lactate OLYCOLYSIS I CIOLZ Foru & Interdebon barca dy motywiod the P Pode that we might opeet FURE 8 illustrata come the synergies and antar Onions between the glycolyik ned the peatore pboqhate pathray. Hetokorak cts in both systems in the one direction Dately from glucome glucone 6 phosphate. Horever in the case of three other easy mel, gluco ·436 Annals Net Work Academy of Sciences pbosphate bobenk aldolase und glyceraldetyrk-3-pboxpbate isomerx the cution procend in głyroly in the direction of Lactate formation m the PP yck the reaction procue Tin the pool dincton mmily toman glococo phaphate formation At the points there an intcrlo king of the metabolc wheels that for them to work in opposite directors Inded if the Pi yek vity Alive on (on sa il gun pushing the irtose phoyihat formed by vyrolysis uk to gli ose-6-phosphat nd, perhaps, lven further lonerd polyacchande synthesis This menul en proceed the FDP axus born in noul 8 or via apothet sequence written us follors (1) G-3-p + S-7-p-E-4p + r-6-p (transaldolas) (2) DHA-P + E4-p SDP (aldolue) (3) SDP-S7 p + 1 (SDP) Sum 2 thac- PF-6-p + The enzyme that atalyzas reaction 3 which we refer to u SDPure bu been dlacovered quite recently It appears to be dely dustnbuted m maxtoorganhos, plants, and venous animal topics An SD Puxe punged from yout was found to have a high degree of specificity The enryme claves SDP to S-7-p and Pi it does not attack S-7p F-o-p or IDP la contrust FDP ak from proach lava that acts only at en kalme PH, SDPure active at PH 70 and may therefore play in mportant role in the reverwal of glycolynos To reapitulate we can not see how the pentome phosphate ycle day pa ticipate in reverung reactions of glucose anabolsun. Tanskelotax and innbolax posh tbc pentore phosphates back to fructose-6-phosphate the sequentul action of innsaldoluk ildolase und SDPure together push ery throc-1-phopbatc und trlovc phoophate back to the berone level The oodation of glucose vn the TCA cycle and via the PP cycle bave imo facton in common Vg++ and thum pyropbosphate which are required for the ovdation of the abeto ucids in the Krebs cyck and for trmketolase action On the other hand there are some striking dimilanties between these two multienzyme ryuteros DPN s required for both glycolyes and ardatne phoxpborylation TP required for the PP cytle Loorganic phosphate bo an ewential ractant in glycoly's and oudative phaspborylaton, but it inbfbuus the direct oodation of glucose-6-phosphate vu the shunt pathway" Spuce po ut only a try sory review of our attempts to study intact excitas tumor cells Ellmg Innme began these investigations of the Cnbtret effect in uscita tumor cells reveral year ago in our laboratory As circumstances did not permit him to complete the rod for publication, I shall dexnbe the caperiments be arried out on the effect of glucose copctatntions on the relative participation of alterative pathways of glacome breakdon a Glocose labeled in C of Copoution was added to ukita tumor cells at high and los concentrations of glucose both to the presence and in the abuence of 2 diotrophenol TAALI shows that at high glucose concentrations the orldawon of a which represents the TCA cycle, a inhibited while Coddation, which represents the firm dep of tbe PP cycle a favored The C/C nto changes from 2 at low glocose concentrations to a nato of 8 at hlgh glucose concentrations. In the second experieL, TABLE 5 the effect of dinitrophenol ·437 Racker and Gatt Glycolysis and Oudative Pathways Tur! ETTICT O GUTOT Cora TINTON OM O Cao LADO a Gurwa T M T wo Cou Total Co, at (1 x) а хто CH1 flacou (6 sko 12 800 6 oso 23 who 2 900 Arata tomo col 20 mg proton) peaded b dan chloodeployphat bota solution 1 A 74 (coatarm 103 codNadu mokes of Pipe ml) natabula Hubur teeter for mm u 39. Co the promo ondactive ghecou, udated abort CHO. vu collected on 02 al 10 N On the center cup ho por 02 nl of 10 N H SOfrom the ode am I the end of the cabaton prod' The peabc Koth ty of CW 1 glucose pred both apamats u 35.000 ct /pok/min Cho thucomo 58 000 a/bok/ ETECT TAS 2 + Domorna O OXIDAD O CALON 1 AO O Orca T AROUND Tmor Cou THCct Le due hidd CH1 clocote Ch 1 pacon + DNP (101) CH 6 came CM 6 flocok + DVP (10 *700 100 3700 HOIDO 8120 9900 2100 1500 Expendatal conditions on RJ Gabon Dinitrophenol has little effect on the vdation of C but dim- Dates the Inhub tion of Coudatlon More frcently llu made extemeve studios Ath limiting factor in the air bohydnte metabolion of Lata tumor cells To punmange bila fodmg (1) Enzyma of glycolyes are present in large excey and do not lunit glucoec utihaton Tho ruchurly revealed by un amly of the odindul azyme of glycolyu., oswell us by mawaring glycolyes to forufied homogenata, which us found to be myenl timas higher thun that of the intact tobaccell (2) Determination of mercellular concentrations of dealne nucleotiden, porn phoxphate, and Intermediates of glycoly a strongly pointed to unor panxc pbocphat u hmt og factor Since ATP levels were high during odde tion, it mus concluded that, either because of compartmenladiatka or beat of the pace of Inhib or the utilitation ITI (or ghore phoquot 110 xu Dexcot under these conditions (3) The limitation of glycolymus ly the intracelhub Pi concentntion abo unduated by a penents on the flect of high concentntons of adkled Inorganic phosphate itxu found wat, both ktobally and memobrcally high cos entrations of borganic pbosphet Amolt glucose upual in luctk ac!

·438 Annals New York Academy of Sciences formation with the result that the per cent l'uteur effect diminishes with incrauung phosphate concentntlom. (4) Tho trumport of inorganx phosphate, therefore apparently limits arbo hydrate Detabolumn of anita tumor celk. As determined by phosphate inayau well us by pe masurement the transport of tnorganic phosphate i promoted by either glycolyus or respination There iko metabolic process Can be used fodendently once their contributions to P uptake are almost additrve In studying the arbohydrate toetabolism of intact cells we have begun with a study of entyme we proceeded to a study of the intracelular concentn uon of intermediate and cofactors and we are sull left with an unsolved problem of permability and active innport References 1 URL RACON Regulatory Niechanon m Curboby dnt Bintabotom III and IV Ip prepunun 2 GATT SKE RACIN Raktory Vechim a Carbohydrat fetabolum I and In proportion 3 KILLIY V TURK KIELLIY 1933 Aspeasc adenonne topbophater or Lover mitochondre J Bal Chem 200-223 4 CLAMI, HG 1979 Obernatoas on the arbohydrat taboleiro of two Brocher J 13.36 S RAO E 1957 Micro and Microcyclo in arbobyenu metalotoo llarney LC Ser 81 143 6 RAR, LEAR SOLLE 1958 Speafic C 1 pooghatak kar Lnxcom 1 6 dupbonphat od nedobaperloee 17 typophute And Brochem Biophys 14 320 7 IRAVOL, CAŁA I GOALUTO 1937 Role of thorpe orthophoupant in herum prophet metabolito Am C D Soc 1.320d 31ty Aber 23 11 turo ·DISEASES OF GLYCOGEN STORAGE WITH SPECIAL REFERENCE TO THE CARDIAC TYPE OF GENERALIZED GLYCOGENOSIS Pal A di Sant Agreke Drvodari, Coorhus Uudy on the Deble drepte, Xa To KY Daca of glycogen storage ire ne disorders due to congenital and at tma familial cross of arbohydrate metabolin lading to abnormal accumulation of glycogen in certain organol the body which us a consequence may become greatly onlarged In moul instances xtend order or the arc el lected to some extent, but the largest accumulation of glycogen ure pt to be found other in the liver or in the hart, so that the dotinct Abdivrops of glycogen doette have been Inditionally distingumbed the beput form and the arduc form. The latter is in reality . geoerilized dycognoos whom maajor dinlaal experts ardlac enlargement and lure of the hout function In addition, other types of duturbances leading to glycogen xong have been recognised in recent times In th bust five yeun momentous advance bave been made in the study of thek dbau These developments have shed an entirely pen light on the study und daugnoof these conditions and have clanfied t a certain artcat the truc Datare of some of them. The chemical studies of Gerty Cort' m particular havo gratly chanfied the fubject and have aborn that m BOTTO imtanca . angle enzyme defect is responsible for the disturbance n curbo hydrate metabollum iod for the abnormal accumulation of glycogen Chemical Background Glycogen a highly brunched polymcchende with a treelike structure Glucosits only building block Biort plucoke units are ) Ded together by +1 uglacodder linkegan to foro chelne In addition there are o-1 blinkuge at brunch points (7102 1) Tho muyme pbosphorylaxe only makes or breaks tbe 1 Hinkages It sydthema fright-chain polyuccharde ll'hen phosphorylaxe ucts in co func thon rith myb-1 4-1 o-trunglucosidax or brunching enzyme the product m the human s dycogen For the soluting of 16-bonds Aul another ennybe, umylo-1 6-glucosklexe or debroching enzyme is required In lycom. deynded by phospborylore the chain lows one glucose rendue uod docose-1 phopbat formed Phospbordlysb Hope as the outermost ther of brunch points of ach molecule a upprouched since phosphorylax an neuther braul por bypup the branch points. The polymcchande tus ootained b aind phoepborylux limit dextrin The debranching enzyme tben removes free glucore from the lunit derinn Glucome and lumit demtrin mind the outermat txt of branch points are the reaction product Pbosphorylax in the prexact of porn pbophato con The ort deonbed the pepo Brorted o put by the Corn Food for Habal Rearch, Babos Hospital, und nut by price from the North Foundat Loth Nee Yart, NY ·440 Annals New York Aademy of Saences 000000 0000000 UTER AA CHES SINNER DRANCHES Fun 1 Model of mento throom molecule Open arck, plecore rende 41 4 binkage black area, rondou m 10 boligon Reproduced by permoon of Aadco Pro uct gun on the polyucchande Acting one after the ther the two ensync degnude the enure glycogen molecule In the digestion mirturo only glucone and glucose 1-phosphate are found Their ato wa marunt of the percentage af branch points in ach glycogen mmple Therefore the spente and altemating use of phoqhorylar and debranchet B an enzymatic method for the study of glycogen structure In body troues glucose 1-phoxpbete u trundarmed to glucose-6-phophate, a reverable reaction medited by the ensyme phosphoghucomutaxe. The liver u un ogans entrusted with the highly mportant tuk of mantamning blood glucose ut normal levels It performa glycogenolyxiu, carential to proper homeostas, by breaking down glycogen through of course glucose-6 phosphate to glucou, wbuch as then made available to the blood uream. This hant recta u mediated by a specific easyme tbe bepatic glucone-opbox phatase, which s aloo found in small amounts in the kidney. The function of glycogenesis, of itplenishing the glycogen stores and of deunanganned blood level of glucose, is arned out by pboqbyorylating glucose to glucore-opbou phate, a reaction medated by the enzyme hesalnak It aun then be inn farted to glycogen through the reactions described in the preceding pangraph. Aade from this bighly specific function of the liver glucose, 10 trelf a meta bolically pert pubstance phospborylated in all normal cells to glucomephosphate through the catalytic actioa of berokinase an entymo ubxquitously present in all times From thu point it may so one of two non, depeodmg on the needs of the body at the moment. It may be transformed to glycogen Lod stored for fature ux or it may be broken down by way of the Euden- Bleyerbof cycle to pytuvate Seranul pathways are open for further procent ing of pyruvic nad again depending on the requirements of the cell at the time.

·di Sant Agoese Diseases of Glycogen Storage 441 Record Types of Diseases of Glycra Story Von Grerte in 1929 dernbed the chancıl and pathological findings in two children bo had died with mueve enbargement of the liver und Hocy duc to deposition of glycogen in there organs He named the condition hepato- Dephromegalle glycogena Chemkaal studies kete pobrequently and out by Scboenhelmer on materal from one of these patients, showing that the glycogen from the ar could not be degraded by minced normal human fiver In 1932 Pompe In Holland and Bachofnod Putscher in Germany todependently described patients dymg in infancy Mth an enormous largement of the bart du to diffuse depodtion of glycogen m the organ. The liver und kidneys et bot significantly affected on these infants By Dalogy with the am of von Gerhe i syndrome, Pompe named this codition cardiomegala glycogena It is recognired from the beginnmg that there wus no pumilanty m the dinaal pleture between what came to be called the bepatic and the ardac types of glycogen storage doenk It was thought for many year, bowever that the metabolk defect auung the expensive accumulation of dycogen wu kimilar in both types The confupon has been heightened ance by the reporting of cures charact med by depouton of glycogen un verous orgins or venous combinations of organu Theme reports have been poorly documented and they have relied primarly on clunial und pathologal findings. It must be liemed that mrphologic changes due to the succumulatbon of glycogen are very umilur even though this polymechande may be quite different in its fructure With few cpcrpoons, ather there are no chemical studies or they were limited to determination of the percentage of glycogen present in the toga mbjected t analyw us compared th pornal finding In very few instances the behavior ol glycogen in the presence of healthy minced liver true wu investigated In 1950 tonna of the arduc type of glycogen konge doreak were reported from this chinue and attenavely Audied from the chmial pathological and chema stud points From the findmg mi ther patients uod a review of the literature it was concluded that the cardiac and bepatic types of glycogedod's were entirely separat disens attributable to different metabolic enton wut suggested by Stetten and reported to our paper that the length of the outer bruscha of glycogen might be abnormal This bypotheds proved to be prophetic in vxw I becquent development. However It only in recent yours and with the application of the techniques developed by Con' that some lyht has been shed on this obscure field To put the cardiac type of generalued glycogenods in the proper perspective the vinow type of denk of glycogen Honge must fint be briefly condered Frve distinct types have been recognized thus fur (TABLE 1) Types L II, III and I are feratu The iplormation availabl onlype y binadequat Type I The type the most ormon, the called bepulk pycogen ronge discou the best doxake of you the of which more than one hus- Ired uno ha e been reported many sort he been seen witbout being regut taad m tbe litenture In the form th liver and at then, the Ldncys are pomanly affected rod urally greatly enlarged. The cbemku structure of the It ·442 Annals New York Academy of Saences TAJLE 1 Dok or Geroon STOLLE Cher Ty Orphed ctna mo Liner ng >100 Normal Absent Abeat Glucose 6.pbopbuto An tinda > Normul ? I Hepatic type Iyon Guate duau) Il Cardens type (genanbod dy caponou) MT Limit dat 04 (Con) Normal Nocol Liver o de, beart IV LV (onbon) Lin Teux caudoth & Abnormal bart outo boncha Aboom long outar branches 1 Normal Abent Debranching my me amylo 1 stocom due Abwent? Brunching mo amyb-14 16 tringtocondes ? Nome! V Dutco typ Juncke cho Orda muner glycogen u normal, but the glucose-6 phoopbatake in the liver and sometimes in the kidneys us abent or reduced. As a consequeext the carbohydnto cycle in the liver s interrupted eod glycogenoływa does not take place in the usual Clinally bypoglycemia, ketons, and hyperlipem la occur and there failure of the normal glycemic response after mlection of cpnephrine Tho latter is a very unportant clinical tent for this type of dreux. Type II The arduc type of geperuired glycogenou's or Poropo dluke LS much less common only about twenty cars that are acceptable u nich bave been doonbed so far Here gun the structure of glycogen la normal, and tho defect leading to the muove accumulation of glycogen throughout the body and having its most important clinical expreadlon in the cardiomeguy la un known All chnical investigations of carbohydnte function and specifically the cplocphone tolerance testere Degative us the glycogedolytic function of the liver is preserved Toy particular form will be further documed later in the paper Type III The fort u chancteosed by a glycogen of abmormal structure with short outer branches due to abuence of amylo-1 6-glucosidase activity (debranching elsyne)" laadmg to glycogen molecule resembling a "humit derton Con, who fint ducovered this type od glycogenous, bus proposed the Dante tout destrinoeth bat pertaps Cordoncise might be a umpler one Specimea from nme an have been analyzed by Con thus far but only one of them has been reported in the clinici hterture Arulable information therefore bounty It wpparently astublished that this condition a fumuhal," that the liver u greatly enlarged, and that muscle nakden of moderate degree may be present Glycemic response to injection of pinephrine Octure, but may be less than bom The heart to apparently not enlarged roentgenio mphlaally or electrocardiographically although this is not known for all cu The pathological findings at autopsy abo ure not available although, on cheo ·Type IV di Sant Agnese Diseases of Glycogen Storage 443 kaalanalye, Pecuneta from tho patents aho bad post mortem exulations sboned glycogen in excesive mounts in the myocardium, u well us in the liver uod in the arate muscle The percentage of the polymcchende in the myoardrum was lower than in the cardac type of glycogen gonge docx Con recorda tbat in som instancusu dagoons of the bepatx type f glycogedoo (von Garte 1 doaure) bad been made chinically but canal maly showed the glycogra to be abnormal and the aux in reulity t belong in this Group The livet uod retkuloendothelul tyttems are offected in the form of the disease and there 11 marked bepati ciriboga nod hepatomegaly Only on care, obnaved at Baba Hapua, New York, NY has been reported in which chemzaal kudlas were done u The structure of the glycogen wus boormal in that this polysacchande had few und acadrely long out a branches it wus thought to be doc to decreased activity of the branching enzyme amylo- 1 4-1 6-thun glucoaldax Tyto v Even lau information cuts concerning TypoV Ooc patient, abo keen at Bubra Hospital, clinically presented u. child two od one hall yaan old with general muscle maten und palatal panlyu Caras umilar to thu ore have been descnbed in the literature, but not studxed chemically Strat muncle on pathological cumination had in appearance kimilar to that of type II and IIL The heart vu uninvolved chnically roentgenographically and electrocardiographically and texts of carbohydrate function were por Chemical rumination of a blopsy peamen of Ante muscle revealed that tho glycogen content wus 13 per cent, but that the fructure of the glycogen u podal Ito worth while to vote again that the patbologra ypatince may be amilar in different types of glycogen xong disaik and that only chemxcal analyes, in addition to the chnical findings, ha aven a clue ut the bank proces Cardiac Glycofer Sloraga Dura At Babes Hop wthree and generalized glycogenous aardiac dycogen storage dog have been obeerved The first two were brother and a star and have been previously reported The tree of the W family la represented in nouu 2 The patient. father was born in Youthern Italy and the mother mus born in the United Suta, bt ho parents had emigrated from southern Italy and were first con Both parents had many mbling, some of whom bad died in Infubcy of unknown ca The first child Lacy ww born in 1922 and delivery WU ponul A few weeks uter orrtå tro parents nored to chey debedor hart bracebby" ind, rome time later the yppant of intamittent cyanos, mare marked during feeding Tbe patient mis admitted at the wo of four mo than a munbund condition The infunt diad De boun after adm.com At autopty the bart mu peatly enlarged On mikroscopic enumetno the arduc muscle bad but later alled a typical lacerat appennt, with much of the myocardal fiber replaced by glycogen In the motions still A iwlable, coidence of glycogen deportion was found in other times ·44 Annals New York Academy of Sciences 9 오 모 YE ir N FALKY PICO ALICY HY HM HO LIVIO CASE MAC POLANE CASI Ο πMAIL RUTECTED TICLE 2 Family treed the family Reproduced by person of Pata dugooers of diffure rabdomycometom of the heart wus made at the time W ben, about ten yan later cardac glycogen storage diese wus recognized u an entity the diagnoses wus corrected. Of the rest aght ablings, tro Anthony and Theren, died at other bospitals at the age of three and one half and four months, respectively of an illness that m all mys resernbled that of the first child The clinical and, in one cure the autopsy records were unfortunately no longer available Las was born to the same mother in 1948 un tult of the tenth pregnancy Pregnancy delivery and early peonatal period cre uneventful At the age of 7 weeks the parent noted a peculiar type of "bavy brathug that attracted their attention beurse it was amilar to that of the 3 chuldren mbo bad died At the age of 2 months the infant begun having spells of cyadosu, cpecially poticable after feeding or ben crying At the age of 3 months the patient wus admitted for investigation. The beart mu found enlarged in all denetet, the wounds weru f poor quality and there sui soft bloning syrohe murmur heard all over the precordium The liver mu felt 3 cm beton the contul margin the ti f the spleen sus palpable. Because of the family bstory ·di Sant Agnese Diseases of Glycogen Storage 445 Fon 3 Root genom altke chart in Low (tabe dance, 2 m) Reproduced by pero al NATO I tentative dugoods of ardiomegalia glycogenkaa was made Cynoms wus present only on feeding in the first few weeks. The aphal beart nuto remained between 130 Add 140 per min The patient deterlorated gradually ha respin tions becaume more rapid and Labored cyndous become cortant He died after 33 day in the hospital A roentgenogrim of the chest 3 days after admbarion showed a plobalar beurt enormooly enlarged in all dameten (NOULE 3) Electrocardiogramare vealed a marka left as deviation, blch is very abnormal lor a patient of thu ye mdedy hovoted T wo in lands II and II ad opborr in land The ST rement u depremd in lands I and II Pleny ante morton chemical investigations are arried out and are all peptive. In particular glucou, pluctor, aod kvaloso tolerance torts et pormal. The glycemic and lactic od rapporto to injection of opinephrine vere Dumoul. At Decropsy the heart so enormously enlarged it weighed 148 gm u compared with the normal 29 go It mus globular and the largement sus fymetin) and pormedy venirebs uitb doproportlomiety od orca, quto typucul m uppearance for the doar The Teatricular albere thichcod and the myocardium paler and furmet than mal VkTocophally the nyucudlum had the typka Wesoch apreur pet (UF 1) each fiber bollon cylindket surtoudded by thin, dexate o toplant, the mucles lymg frre in the enter duct the masne infiltration of glycogen The change in Aruture us ko grata to male recognition of myocardal ta dificult Stnking chango o alo found in theletal muscle. The areare dometer ·446 Annals New York Academy of Sciences For 4 La beart much, born central clear are lo Aber and periphol Arcople Naca are found in conta o fbar AD fibers are involved (alled bactwork 4ppartaca) Hemelorylus and con X100 Reproduced by person of Panay of the fibers wu increased from 2 to 5 times moderately affected one bad rows of vacuoles lying between fibrilac, bile in the larger fibes the entert cytoplasm appared replaced by glycogen The amount of the polynccharide vaned greatly in different muscles. In virtually all other times, including Horooth muscle, liver and other glycogen we present in large amounts Results of post-mortan chemical studies showed that the glycogen content wus highest in the hart (98 per cent) but not much lower in the tongue (6 6 per ceat) pou muscle (6.3 per cent) and liver (71 por cent) Normal value for glycogen content us given by len Creveld vary from 0 07 per cent for the myoordium to 0 103 per cent for the bepatic tissues After other amples of the same too had been kept for 24 hours in the incubator at 375 C chemical analyses revealed that antoglycogepolyes had taken place o all inatances, although the degree vaned in the different organs The third patient we observed mus D B the third child of the B furnily A previous obling had raffered from the cardiac type of glycogen Honge doense and was reported in the literature She had died at the age of five und one half months The second child in the family mu normal us wus the fourth sbling The Parent were both of Englash extraction, born to the United Stata, und sot rdated DB mu bom at our bopital and followed from birth which afforded in Un al opportunity to study the development of this doreak On phynoul ruanoatlon shortly after delivery the child appeared quite normal, bad a good macular tone, but was found to have a hanh murmur Roentgenogram of the chat on the fourth day of life shoned the heart shadow to be patly enlarged lo al duidelor However the electrocardiogram, done on the orth day wus comdora non ·47 di Sant Agnese Diseases of Glycogen Storage Tuu 2 Post lomo CONCAL Stupa D. TO Chet 14 de ta Depndebo olm Actory (c) 64 (ra) Hart Lner Addoctor Toro Rectos Draphy Extoty 71 60 66 12 Yormal III lant ||1||$98 Karol Bruin 04 Palermed by Gerty Con The baby appared to act and grow normally and be had no difficulty feed. ing until the age of 6-cek, when he began to bare marked respiratory dlatrem. He had aincultatory nila all over the pulpookc areas and the liver wu mark edly enlarged He is considered to be in Letture. After adminion be Improved conderably Digitalu sus gren The beart was markedly a Larged co Y my cod the dectrocardiogun showed laverion of the Tiiva und daprendon of the ST segment in several landı Tbe changes continued progresively Blany chemlal tests were performed and were all within Dormal limits. In particular the glycemic response to pinephrine inyection was quito normal Subacquent epbodes occurred chancterized by cyrunosla, respiratory datre, and enlargement of the liver feeding beaume difficult the pathent became programerely worse and he died at the qe of 3 months At Decropsy the findings were ordilar to those of the other patient There wu an enormowly enlarged, globular bart Dicroscopucally the appearunce of the myocardiom wu quite typical of this doex and there mu mkroncopx evidence of deposition of glycogen in all tissues of the body The iperunce of the top and in particular of the striate muscke van fn all man umilar t that of other patents Al too wore frozen 90 min after death and were kept on yeniloc adone by ur br dry ke I Certy Cori in St. Louis, Jio ho performed all of the chemical mlyne. The results are luted in TABLE 2. AD tas unvestigated contained a large arboant of Ulycogen The content of this polyuccharide us httle bigher in the mayoardlum tben in most the ith the exception of the top tie but the fures are condy compunable in the live ther. There were 650 mg of free glucose per cat in the bopatic theex op amlyla, od moother 150 mg fortoed after focubation of the liver trauc for 3 boars at 30°C. In the adductor urlate macke that there wac 273 mg, of lactic acid per 100 gm. There realts indlat that in both liver and miracle pycogen could break dos, quite in keeping with pola studier Glucose-6-pbocplutax activity a determoed in both fiver ud koey that und wu found t be normal Structure analyses of dycogen ·449 Annals New York Academy of Sciences Rith the enzymats Drethod ung alternately phosphorylase and the de branching myme amylo-1 6-glucosidak. The glycogen gructure vos con udcred Dormal. Specimens from arm smili wao bave been suloquently anatyred by Gerty Con and havo yielded similar finding Condessons It is evident, therefore, that walled glycogen kong discuse of the heart (Type II) u 1 spante and distinct entity among the diseases of glycogen stor 4ge. It o in reality a generalized disorder due to a metabolic error affecting arbohydnto metabolim throughout the body and not only in the beurt. Car diac enlargements its major chinal proroon and the aux of death. The no-alled aardac type of glycogen gorge is incompatible with life for more than a few months, during which carduc function continuously regresses The most plausible crplanations that of a mechanical dath produced by an accumulation of glycogen in the myocardial fibers to point where their contraction impeded and effective arduc action no longer It s interesting m the respect to note that in the glycogen at nago dicas alled hmit dextndows or Cari dirase, our Type M, charactensed by an abpormal glycogen Bructure analyes by Con in tho ako" show an ice of glycogen in the myocardrum, u well as in the arute muscle The levels in the crumined peamem have been less than 5 per cent of wet sexght and, insofar us known, patients have dether been in verious trouble chnally because of their arduc function, por have their electrocardiograms or roentgenograma been aboormal Apparently the percentage of glycogen in the myocardium must be above this level to cause symptoms ben it reaches 7 per cent or above it becomes compatible with ardac function What metabolic defect lads to th genenalized and abnormal glycogen ac cumulation in the myocardnim ud throughout the body in the carduc typo of storage discuse has not been uncertained but the Rudles we have made thus far shed some bght, in a negative way on the obscurt subject On clinical grounds we knew the glycogenolytic function of the liver ww preserved and now we have the masurement of glocore-6-phospbatore showing it » Dormal. The structure of glycogen also pormal in the type of discus There 11 raron to believe that glycogen can be uulued by the body in there cines because of a preserved capedty in the live end the muscle to break domp thu polymacchande and because of the absence of demonstrable ac cumulation of intermediate products of carbohydrate metabollen, mich u Pyruvate or lactate, or of uny evidence pointing tomard the combixtion of large imounts of fat The carduc type of glycogen storage docare ia hereditary ind presumably It is inmitted u 1 receve It nure, us shown by the fact we have seen only 3 area of this condition in two familia at Babios Hompital in 37 your and in more than 3000 rutopea, and only 20 ases have been recorded in the Kenture, although many more have been xen, but not reported Criters an be listed us follows for establement of this duguous (1) maried m burgement of the bart (2) dath #ithin the fint year of life due to haut ·di Sant Agnese Diseases of Glycogen Storage 419 failure (3) typical lucem od appannce of histologau salons of the myoardrum rcalting from mopshe deposition of stored material in all ardiac fiber and (4) post mortem chemial studies demonstratmg normaal glycoger stucture and preserved glycogemolyes m the liver und n tbe strate muscle In the differentml dugoni other conditions gring rise to andlec hypot trophy m infancy should be considered arch w endocardul fibroclustoo, virul myocarditu, and aberrat origin of the coronary artery Electroardro phxc findings rugrative of myocardal damage can be seen in many of these conditions and the oentgenognphle apparence Dot Concheye. A biopsy of the triate mixclo showing th batological and histochemalence of Ictave glycogen deportion th the chanctastic degengnion of muscle fibers that accompanies it is highly suggestive of the doux. It should also be kept in mind that there are many other circumstanca in which are glycogen - present in the myocardrum, but la potty and not generalized dutnbution Rhabdomyora o the heart, soalled animegala dycogenic circumscripta, and cases of pudden death in which acetve but localized accumulation of glycogen a found on the myocardrum after denth are among the conditions There is evidence that, ben pyocardul cella are destroyed by a vanety of yeata, infiltration with dycogen octun Th. s obviously due to a different mecbantem and should not be confined th th unknown metabolic defect that lands to AIIVT glycogen storage in all tous of the body mladog the myocardium In concluson the dress of glycogen xong, in particular thome of the arduc type, constituto a very obscure field in which a funt afirmer of light has been pred by the collaboration of the clucan, the pathologist, the ardmboght and promourly the chemist DIuch more orl romains to be done and, if further druncar ure to be made in the confused bold they will be due to continued cooperation mong disciphna In particular applaatoo by the chemist of his specialized Epowledge and techniques will undoubtedly prove highly rewuding Roannces AX CHILD S 1939 Glycopa Vadece 101 2 DIAN HH&DH ANHY 1911 Ch com a AD J Dau Chekten 0793 JD SAWT20IE PADI ADALAH Valon 1930 Olycogen wong du ct of the bart II Cntalitetbe htanut Patra 607 Coel GT 1954 Gh coga fracture and you defoong in scopo storen di Harry Lecuo Ser 145 Son mı E 1929 Fipato napkro pagal dycoona (Chloggodalnak bat da Lobe od vel Betr putbol Aont en Puubod at 497 6 ScoutR 1929 Uber abe egenartig Storing dan Kotlebuttato che 2 pkyeol Chan 10:18 7 Pan, IC 1912 o Waube by peteropbor u bet art Ned T skoder Cake TI 301 8 Detro 1911 1912 Zon lencber Bu der Gk lopen pertukat iGh Lomon) Zwerber m722 OPUT 1912 let aspeteree Chlamydinnlleu de loin the VINO chetgenkaBeitr putbol Aant to htbol 10m SAYLIIDII Auror [ HTHIRADI 1930 Gha o ponto the bort I Report at com ho #bdomalud rubo bgcoca Podatno #02 1 ( 10 ·452 Annals New York Academy of Sciences SCHE 1 PUHDU EDO CO+Foss REXDOTI Ox OID4DTE VIDCHAT CO, + prunito + TPNH Amulete + TPN 11 CO + pbombopunut + GDP Bulaceut + GTP + konty 8 COA dirty, etc Pyrinta fatty acids Sons 2 PounL ROLE OF CO. FDIATDA DX TE FOKADOX O PHOEDOPTIUVATI NON PEUTATE CO, + TPNH + pronat milato DPN+ omelecetate + DPNH GTP CO + pompbopgrunto + GDP DPN It s pomable to vulco a role for the Corination reactions on tho forma tion of pboxpbopytuvate from pyruvate, as shown to CHIHE 2 In thr proposed sequence pyruvate converted to pbop bopyruvato by the combined action of the two Corfiong enzyma, although e number of other enzymes we abo required to apply TPNH and GTP to remove DPNH, and to andize malate to ovulacetato The pet raction cacatuly the conversion of Pyruvato and GTP to phopbopyruvate at the cipense of the oxdation of TPNH Pyruvate + GTP TPNH + K 0 (3) pbop bopyrunte + GDP + TPN+ + HO Thos reaction sequence provides i mechanion for the several of the pyruvic Hnaxe auction pboophopyruvato + ATP = pyruvate + ATP Raction 4 hos been shown to be reversible, but BME 2 TA proposed a more fanble pathway for phorphopyruvate formation on thermodynamic grounde! The equlibrium constant of REACTION 4 has far to the right while the equilibaum of Rection 3 should favor pbosphopyrunate synthes Although GTP is believed to be the pbosphate dopor In CISE 2 it can be ·Utter CO, Firation 453 farmed from ATP and GDP by the muchonde diphosphalfuxe taction ATP + GDP #ADP + GTP (5) The present domingon will attempt to caamine the aulable data with Occaporal reference to CH 2 to ke If the concept embodiad In CoE 2 by any vallity It will be a particular interest to consider y Doo true indepeodently Data obtained by the one of botoprally labeled rubetritos support the beld that CO, tuation is tovolved in the formation of liver glycogen from substratas at the level of lactate ar pyrınte It has been wel establwhed that isotoprally labeled , bancarported into the 3 und 4 portion of the hero molety of hver glycogen In addition, Topper and Hasting ind Shreeve" thowed that, m the convention of 2-Pyrunte-Ct glycogen by Irver whices, the detribution on glycogen that expected if the pathway mvolves a symmetral intermediat much us fumante, u shown in WHAT 3 Соттерол SIO 3 CH, CO COOH TO G TOOLK CH CO 01 CO. COOH CHCE DO OE CULCDE ODOH 1 MOH CI CO COOH CHCA CUOL CCCCCC If yyrunto labeled u indicated a converted to meet by CO, fication, the label will be nodomued between the two Doncarboxylic carbon atoma by fumarus, with the result that th phopbopyruvate lormed rubacquendy will bour 2 od 3 labeling, and glycogen herom a 1 2, 5 and 6 Labeling 11 Pyruvato a converted directly to pboxpbopyruvate without pandog through rymmetna intermedute, phopbopyruvate will be labeled only in the 2 Poation and glycogen bemoe only in the 2 und 5 pontinos The actual epentinental results 1 showed that the glycogen wu labeled almost equally in the 1 2,5 od 6 poutsom upporting the fint of these two pathway for phopbopyronte sother Unfortunately u often the car to sotopic exponents, the rezults ain abo be interpreted us the result of cichango Tactions that do not necessarily reflect Det synthemu. ID CH 3 if phot pbopyruvate = formed directly from pyruvato (dotted Imo) qallibration of the phophopytuvat with the daarbarylac ac an ladt trundiALO of the label m the phospbopyruvate and rubeequent formato a dycogen with the observed Labeling W bave investigated the pondble relationship of the Corfinog ractions to pboqphopyrunt formation in 1 mer hat different whlon Pennington ·454 Annals New York Academy of Saences TAU 1 FO or of ITO TOTIUTATI nor Por te Incabatt Moplaya Machine 888. 0 0 25 0 47 014 0 0 13 0 25 000 Incubation pont en contum locore 6-P dehydrogenas, make myne, daphonx, PEP arboylu, od make deb; drogtunk, u vel phacoee 6 P TPN DPT ITP N Pyruvate loc ind AaHCO, Total volume - 4 ml temperatur, 37 € TALE 2 POHOIDOTATE FONTDOW BY DARXOXTLC LCD Pane humb Moplogrunt 0 41 0 08 0 16 Completo TPN DPN ITP Pyruvat OOOOO OO000 Conditions and components on the me u tbon m TANI 1 Tum, 60 min and I first tried to accomplob the formation of phosphopyruvato from pyruvato by an in turo syetem composed of the vinou enzymes and gabentas that had been suggested us taking part in wh 1 sequence of reactions, Those attempts have been succesful, but l bo touched on only very bnefly As show I TABLE 1 when malxc enryme und PEP carbotylinak ere incubated with pyrnate, CO, phosphate donar and the reqwred budury enzyme and cofactors, phoxphopyrumte iod phosphoglycente, are famed TABLE 2 nbow that the various cofactors and substrates are all necemary for the raction to proceed There speriments demonstrate that it is posible to TABLE 3 EYAL Lung T RA TU T. center Laura OS S2 93 detal pack Brin Haurt Kidney Lle 0 02 0 14 0 21 29 14 1J Acth ty - moto /10 mg protein Ang were and ot astrid o scetose por der of the PTI kod Deyme a unyed pectropbotometroally to C PEP cambory Line 135C by Chachango wtb lootato ·Utter CO, Firation 155 link together purified muymes to carry out the doutorytec pathway of phos phopytuvate formation. Suated in other tone, the patha y has been shorn to be fcashle in a Linetic sro. The foregoing perlments were performed with treyma obuted from a variety of sources If the dicuborylic ad patbs y has any abdity all there enrgmos most be present in reasonabl uodu m . angle tasue We vete fatacted particularly in the tro Cung enzyme, unce other evidence anılable to fodicate that the remaining MIYToe hare e ude dotnbauon and were probably present in all the too tarted harordingly Hm und I hare menored the amounts of malc cozyme and PEP arborylimuse m woos Darde TALE 3 shows the relati e Dounts of these myma metrict of Ketooc powdas o fire nt tissues. Pyruvic imas bus aboo been included, oct It consultata a poorable alternate route for pboephopyrnate syotben. The most significant finding keem to be these (1) PEP carborylimuse is found in liver and Laney The small amounts found in the other to approach the wengin ty of the 105 method, at though the value for haurt and brain may be sgnificant (2) SIulic enzyme has alality uniform danbulion m the trues tested, with the exception of a lower value for skeletal muscle Even in de th value I denibant although in arler Audies umg ı defferent umy procedurc, malcentyme had not been detected in Dgnificant amounts (3) Pyruvic kinux is found in all the examined, but there be large variation with keletal muscle containing about forty the us much of the LEYTDC usb containod n liver The qunutative estimation of enryma i topues has many pitfall, and the values obtained may vary with much factors us the handling and etc tina of tbe box, the muy procedure, the presence of interfering enzymes, and the species of animal croployed A venety of procedures has been the In the determination of these three earymas innt, nabbat, and cold en toe and ultboogh the valua obtained very sooxcwbat depending on the above factors, la ll chro the general pattoms that shows in TANE 3 I believe that these relationships will prove to be keptally correct with the following species differences the CO ng corymo are considerably higher on specific uctivity bugs in ustan liver darparty in pyrum look content of nerous tlustes to even more marked in rabbit and chickon the than a shom a for attoos I TABLE 3 How do these dostnbution studies are with the previously outlined roks of the Corfination rouctions and ith other Loor ledge of the blochemustry f the Furloos tres The ability of the hver to form dycoçon from Lactate pyturats, und certain imuno ulds a well knon, and there babo consident cvidence that the widney co produce plycogen glucome from dmilar amb Kata o If thytoren s formed via the daarborylic acid pathway these tbruas should be di rapplied to mafie enigme 1od PEP arbory K. Indeed, we find that there are the best uod pertape only sources of PEP arborykimase and that mala cosya baboo proxent la coordenah amounts in these toas. Converxty the pytunk kthere in there to be the lowest of Jl those camined ·456 Annals New York Academy of Sciences It might be polnted out that phosphopyrunite solbesia by a pathway Auch u the dicarborylic acid route would be largely Degated by the presto of an act of pyruvic Lucas, us thus enzyme would reconvert phospho Pyruvate to pynimito in the presence of ADP and 2 mechant for the rtgeneration of ADP from ATP There is little endence that skeletal muscle or brain can form glycogen at any appreciable nate from rubatanco richa Pyrovnic or lactate Theke tumoules contain little or no CEP aarborykiner, but are the highest in pyruvic kuase conteat CO, finton may still play . role in these twia in the synthesis of the dicubotylic acids needed for the operation of the Krebs ycle, unce malic entyme a present in both cukor In a qualitative way then, the results of the engyme distribution studies in the four ma just mentioned support the hypothes that CO, fixation plays a part in głyr ogen tyntheos from 3 carbon acid, The Rution with regard to the heart less mtbructory On the basis of the corymatic #wes ono would conclude that tho conversion of 3-carbon acda to glycogen by the dicarboxylic acid pathway must be slow or perhaps beat Other types of cvidence fuggest that lutate may be averted to glycogen in heart although this point cannot be regarded u settled The heart has a substantm glycogen content and utilizes lactate readily as a rubetrate ObecTvrtom that lactate could krve us major fubatrate for bolated heart preparations, meanwhile mantaning th glycogen content, led to fuggestions that lactate might be converted to glycogen As we made dear in the ongical reports, the results can be interpreted equally well ut qunng action of lactate on glycogen and annot be consxdered us cooclturve evidence for glycogen synthesis from bactate Stadie dal showed that slice from at heart could form glycogen from glucose but they were umble to demonstrate glycogen formation with lactate pyruvato or Llanine u substratos Pertupe the most Ribstantial evidence that heart may form glycogen from 3-aarbon PTECTYOTS B provided by the experimento of Lorbet o d' on the incorporation of CHOy into the glycogen of the do lated at heart when incubated with glucose add la tate Theke Torten foued that heart glycogen acquired a pubstantial labeling under these cir cumstance in relatively short time Therr caults Riggest that the solated bart hus the apaty to synthene glycogen by the diabotylic acid pethaay by reveral of the pyruva Linuke reaction a by both mecbanuma It should be noted, however that it a porable to suggest mechanics by which CO, could enter glycogen by exchange reactions that do not lad to set tynthes of glycogen It s unlikely that a peces difference be involved in the apperent contradoc on between the corymstic cod partido esperiments are as dos chicken and at bart preparations all gove tenthally the same results on entymetic goden At present the question of glycogen formation from 3-curboa mads in baart mot be regarded as unsettled. However made fryme be pretent in all bart tria tested in relatively high amount therefore, CO, fiation may play i roke ta tho tosue in the formation of the Faabon dlarboxylic uckdos needed for oddatire ractions The intracellular doinbution of the me three encyna bol some interest ·Utter CO, Firation 157 T + I DATILU DODATION CON VID 3 Pernom IHOX TIO CL txt bu MP ru வ Nitor bondine IKTORIDA Sepetontut + Frictloolllons of chichen I or homogenata cota 025 1 rok Fractionation Hudies of boulogenates of hxckniner by the walceatndagul technique ure suomarloed in TABLE + As indi ated maluc enzyme bahor completely located in the supernatant fra twn hal PEP arborykinase u found mainly in the mitochondral in bon all but yuble amount of PEP carboxyhinax appears in the pipernatant probably because of mitocboodrial destruction during the handling pro edure I'yturk kinan uppears to be distributed between the mokrosomal od apermutunt fruton a mall amount is abo found associated th the mitochoodria, probably us contumimnt. According to these findings, the general relationship between the enzymbo of the roftochondrion and the cytoplum might be represented as shown in SCULE + According to this hypotbess the converpo of pytuvate to phon SCIDUL 4 MOCIOLLON RELATTONIAM AD Crorli Cytoplen Litocoon onlacrtat PEP PEP paco, pls copen 1 Ponut mate acet; 19 cat TPVEL, CO Dilat phopytuvate would involve both the cytopluck und mitocbaodrtal clements af hver cells, with th pyturitelo-malat cooverloo occurning in the cytoplan und the further corvetulon of malate t phosphopyruvut occurring within or on the mitochoodnon I'urthermort mitochondita sbould be able to form phosphopyruvate from Abstances that yield oulacetat Meny yan ago Kaklar showed that kidney particulates ould forto phoopbopyruvat from malate Bartkeyte and Bandunki und Lipoann" bave shown that liver mitochondra 10 form phowphopynuate from daarborylic ackis, uod nc ako have found this t be trae. In view of the uncertainties concerning the powable formation of pboxphopyrnute in hart, it would be of interest to see heart mitochoodrla coul ·458 Annals New York Academy of Saences ako form phoxpbopytuvate from malete. Thus fu our efforts in this direction have beco completely una codul Up to this pomt the comuderation of Corfining reactions in phospbopytunte uod glycogar synthew has been limited to a decurion of qualitatire gode lle Dow have attopted to secure more qmoutative information in the ante od one time, chicken liver Eym of bormonal dietary and other factors are neglected the following information is required in order to determine whether the daarborylic and pathways efficiently active to account for pbospbopynt vite and glycogen synthede (1) the rate of glycogen synthesis (2) the con centration of the enzyma involved (3) the relationship between mbstrate concentration and reaction velociues of these enryma end (4) the 1x 140 concentration of the substnte or abstrata As will be readily reen, it difficult to find u metabolic pathway for khach the type of detailed quantitative Information sivulable we do not have all of these data for the dicarboxylic nad pathway Homever it may be of interest to present briefly the available Information, us it permits certain tentative conclusions coocerning the route of phosphopyruvate sauber's, and abo because I lustrative of a type of problem that will appear more frequently in brochetokaal studio us the quanti utive evaluation of potentul pathways becomes necesary The fint question concerns the nate of dytogen synthes Lven the quch tion cannot be answered with certainty in typical esperimento m which fusted nu are given sodium lactate the amount of dycogen land down in two-bour persod s equivalent to partaps 1 mmole of those pet minute pet Tam of liver Tho nato in live slices s slovo perbapo about 0 25 pmole of trmose per minute per gram. Thex valdos ao be considered only as rough apprauations Ance they are alculated from angle tune interval and po.co the dycogen may not all anse from lactate The concentrations of pyruvic kinax, mali ediyme, and PEP carboryturare mchiden liver are born in TABLE S with the resulta exprawed us moles of activity per minute per mm of live at 37 C The second column from the nght shots the man of the deaminations on series of 10 chuchers Far pnculal raons each of the three enzyme nu mauared in the opponto direction from phonphopytuvate rynthes, nod it mus necentary to alculate the nate in the direction of phosphopyrivate synthems Thos hus accomplothed by the uk onthos of the velocities of the formrd ed back reactions obtained TAILI 5 ACTIVITI Or Eu Covard M ProOnLTY T Sn (Cm LIT) Hnute Eo pihaptent takut Tyrux Linux slabe my PEP arbor, Linas lytun te formato TTVH formation 010, nt lucttat 7 2 30 10 9 0 0 1 35 25 5 Activitas corrected to 3T C | Calcolated from pre os column una nuo od forvard und buchuud oatla ·Utter CO Fixation 459 from eperiments with penfied enzymes Illustrate Pyrinc krake 113 determined by a spectrophotometna isay u shond in REACTIONI 6 and 7 pborphopyruvate + Adry Line Private + ATP (6) har det droix pyrzte + DPH lactat + DP + (7) For a number of reason it is not posible to mourut LLACTIOX 6 m the reverse direction in crude ertracts Therefore the ratio of the initul forward und bach velocities of this reaction nu measured using bichly punfied provt Kirk John McCuato come out amar eperiments in our laboratory and found that the ntb of the fornard and back velocities about 1+0 at pH 74 despite the fact that the K of the reaction at the aume PH s approd matdy 6000 Similar experiments are came out with makc entydde and PEP arboxylinase to obtain the ratio of the mutal velociter mn the two direction. These ratios were then atuued to akulate the values abonn in the nght hand column of TABLE 5 Thex alucs represent the mum um attainable velocitlos of the three enzymes in the direction of popphopyruvate yother under optimal cooditions The valua shown in LALE Surt calculated the bow of ma mom initial velocitos under optimal conditions of nulstnate concentrabon. Ix ato the conditions muy be lewe favorable and the actual relocatio M probably be len than those sbor L The values shown in TABLE 5 are abject to corderable error brut probably establish the general order of magnitude. Even woning for the probable error, it would yppar that the pyram kimx bn birer cannot account for an uppreciable part of the phosphopyruvate synthou o that those. There is posebly cough malic entyme to account for phopbopyruvate synthese altbough the manmum rate attuable, 1.55 winola, fality close to the atimated nate required (1 0 xmole) The concentration PEP arbory Linux Appean to be in considerable cices The entymath yeloatlos documed above are mermum fnitul velodties the actuel rates ik ert will depend upon the concentration of Kabanate ay nilable and on other factors. The relatbocahip of reaction relocity and wabunto Concentration is reflected in the Darchedis-Jenten constant K. There Tulus for the enzyl as under decision are avea I TABLE 6, Indur u they ure Kalabte Ooc of the sport endng pools in the very high K, for pyruvate of pyravic kinase (9 X 10 ) Thla value may be contrusted th the K d pyruvate for mafic eryme bich ks very much long (3 x 10+ 1) and probably much more in accord with plynological concentntions of sytuvate. The K, for CO, for sale caryme is not knor a with any perechiloo, but epal meatı given by Ochoa" und sogy of our ona data indacate that the K. La quite high. I tho to true It porn forradable obstacot the diarborylic udd pathway unless there costs come unrecognierd mechanw far acthruting or concentrating Co, at the ste of coryne action. It is intercating to note that the ane sort of dilemme mbu in the photosynthetic tothon oc with nbulome dupbosphate for which the K. for CO, with the purified ·460 Annals New York Academy of Sciences TAL 6 E VALUD TO SOTTRATOS IXTOLYO N PorrorIT IT ST Than Duy Sabanata Pytryk Lour abc entymet Pravat ITP Punto TPNIL 9 X 10- 8 X 10 J X 10- Yaylor 6 X 10 ) Voy bor Low PET ar mylimut GTP Qalacetate Ponged from abbat marche Purified from Arun hyer is very high When the natunlly occurring concentratica3 of chrymc ere condered this reaction aun bardly account for the nate of photosynthetx CO, fixation The tentative conclusions that may be drawn concerning the role of the COF fixing reactiom aared out by malk enzyme and PEP arborykinax m animal oud seem to me to be us follors marc enzyme has a very wide databauon In anual tras and potentially can participate in the caridative reaction of there there by supplying decarboty Lx uches. In addhtion, in liver and Lidney there u evidence to suggest that malie czyme and PEP cuboxylinan foron part of a mechange for the synthes of phosphopyruvate from pyruvate during glycogeneals Quantitative studies of the unmounts of enzymes present and their kinetic properties are not sufficiently detailed to permit a definitive answer to the probleto of bether phosphopyrmte formation by a pathway involving Co, fizton's the major route of synthesis References 1 OCIOS IH WALL & A KOOK 19:13 J Biol Chen 1741 979 2 VERA SALLO, J B&S OOTGA 1950 Bal doo 167 319 2 VERA SALLO, JB, 8 Ocra & PJ Om Biol Choro 107 863 3 UTIL, MFHH000 1916 J Bal Chan 1h1 455 4 UTO, TAK KOLLARI 1934 J Biol Chem 07 787 421 4b Umn, MFK Kowa&I A Roe 1991 J Biol Chem 207 203 5 K., HA 1951 BN Jabon Hoplum Hop M: 19 6 LAO HAIJA Zoro 1945 J Boi ben 18 343 7 KOLKARL K RJ PONDOTO & H'F UT 1957 J Biol Chem 28 10:59 & Buro, P & K JOULD 1934 J Bal Cheon 110: 657 An Thoop NON LIRON & V Loam 1915 J Biol Chem 111 475 96 SOAT GH FOLL, Y Lom&H Woo 1919 J Bol Chem. IT71 679 10 Torta, 1 JLA BHUTDOS 1919 J Bal Chep 11 1255 11 SAYL, HB 1982 J Bal Cheo 151 12 ROL, JAKAENT 1941 J Biol. Cbcm 1401 747 13 ROLA 1912 40 J Phynot 3 95 14 RIKO RW 1913 Am J Phyri 16: 776 15 Trun, R A 1944 Am Physal 141 662 16 E CLFG WiFi Hg 1005 Court J Expul Phyd M47 17 SOL E L Law & HB RouteY 1937 Am J Phyto 119 404 18 SLADIL, V CNH Tour & Van 1947 Bol Chem 17U 419 19 LO, Y A HONG KAO Nin 1913 J Bal Chem in 617 20 KALOKAL, HV 1919 Biochem J : 631.

·Utter CO, Fixation
461
21 BAITIT 1 1951 Brocton J 64 387
216 Burn 1955 1: U Biobem. J & sss
22.
BLAIRST Lorx 194 B. Chcom 011 741
2. So I DA IGA F1 ) ) Bla&AB Hut
D. 1941 J Bal Core 100 171
24 B, SIŁEG HOLID 1937 Brocheo J 119
25. OCOA, S 1950 ITbc Ene 12 929 J B Seper & II Tad, Lda
tandemke Preu de York Y
2 Renn, E 1957 Irch Brocbco Brophy 0 300

·ENERGY-GENERATING SYSTEJIS Santuto Grbolla Umrly Kasu ka Soka Koca CHy ka It is generally accepted that cardac hypertrophy and failure occur as the repult of factor connected with an increased loud to the muscle and that the hypertropb fuling muscle a befficient This, borever a ilograal dace, by Incanang its sure the organ pandotically increase its difficults for more efficient orygenation. The heart muscle, although m general in acrobac organ, falrly well endowed with dycolytic miyma. I wonder if the book m Are when the chemoglobin erpply los cred is the primary delect and if it Itsults in adaptation to a more acrobx condition at the cell level with a corresponding increase lo glycolytic enzyme I wonder also aboat tbc pot albilty that the control mechanum for glycogen Honige on the heart, under normal and abnormal cooditions, lada to ardhuc fulure The latter problems of particular interest in vxen of the recent discovery and Identificatoa of adenytphosphate anhydride us the co-factor for activa tion of liver phosphorylasel od of the ser pathnay for glycogen synthem from orld do dipboopbate ducone (UDPG) I it powłble that dycogen stored by the UDPG ensymo rysted, that glycolyw proceeds physiologically under the control action of glucagon, und that the phosphorylas become muurally activated by adrenaline as an emergency mechanism? Is glycogen in the muscle len dynamic tban dycogen in the liver? What about heart glycogen doen glycogen mfluence tirsae metabolism and thus contributes more than just a storage function? Some of these quesuon bave been uled prevlowly and, brocbembally speaking, there does seem to be a marked dif ference between muscle and liva phoxphorylare however definite comer x yet to be given For many year there have been reports in the literature on Donorthodox nucleotides, the ti di Lod monophosphata of denonne (ATP ADP and AJIP) for comple the ATP type II of Bang or the adenine dinuclconde a Kellog and Bleyerhoff that appears to resemble if not to be adcatkal with the co-factor for fermentation of Ohlmeyor! Are these colectons, or almlar one, perhaps the baart nuclcoude H, important to ardiac muscle or to its regulation, whether or not at the carbohydntc level? In regard to tha, in Initial ATP sputting in male contruction has been challenged recently Honever the direct transfer of pboxpbate from 1 3-diphospboglycerute to creatine to form phosphocreatme without the intermedato action of an adenine pucleotide has been abonn recently Perhaps, since the enzyme that produces the reactlon 1 S-dipboxpboglycerute Creatine phosphate kems to be backing in bart muck, thu organ khonkd be used to tast hether ATP split intally during contactbon Heart muscle has much lete phosphocreatine and ATP then hus skeletal muscle and would therefore, provide bettar erperimental modd It should be remembered that tbare was splitting of an unidentified organic phosphate durog a doghe music twitch, although 462 ·Grisolia Energ-Generating Systems 463 the ATP cod creatine pboplate concentrato did not change Therefore the 1 3-diphosphogłyterte creatme phosphate section, or a umbr ooc may interfere Kriowly with th ATP and ruume pbogphate balance studies Sunce kome derangements of arduc function appar to be Initially related to metabolic, rather than to roccbaokd deats us in berleben und in bypo thyroxmon, b it possible that there are more inital metabolk defects in the ardac mucke, both before and at fular than we are inre of it pretent? In my opinlon me should not expect to find early dramatic changes, at leust in blochemlal panmeter, in conditions whxb develop gradually I am more that a chalinuou, for example a 5 per mi, change in metabolc regulation would have a very Ku diect in the long run bon ever it is aceptional to be able to measure blochemical reactions in crude emples within a 5-pa cent Dott Thomine therapy has been recommended on the bus of better response of the partially thlumme deficient beart to carduc stycosida in the presence of additional thamine" Catanly there are some clinical cardiopathic labeled Wiopathic which occur Dostly m young o mddle-aged people there patients, although they are without apparent mechanical defect, eventually display cardiac dilatation and fulure Pahips a further indiation of individual inceptbülty the apparent fundamental defect in alcohol cardiopathy add, for that matter the extremde vanability of the Hver i rekisting curbows whether due to alcobal or other causes The hormone thyromne appear to influence both glycolyes ad respiration, and the interdependence of these procentas kell known (Puter and Crabtree effect) Therefore, the upcoupling eats of thyrodne upon otudative phosphoryla tion, the Interern recent observation that the sching of mitocboodra produced by thu hormones dependent on the redor state of the mitochondria (doce under Ancoba coodit or in the presence a cyanide there no effect) the powbility of mong the observation experimental teuing model for permeability effects of despitalos, and the many obervato on Inhubitory ofects of thryo use on dehydrogues, transhydrogenase, crenthe klack, etc will pot be documed here. Recently it has been obeerved that the bormone thyroxine inhibiu completely ucyl phorphatax," un entyme that, among other substrate, an dephophorylate 1 1d pborphoglycenate (1 3-DPGA) Recent work in our laborztory with highly purified bart and bran arbetyl phoghatak o appen to mdate that arbamyl pbopketak end ecyl peoppbatax are the Me rryme." Thys enryme which is probably the mallest thus far recorded (DIT - 19,000 or less) may be important to both carbohydnt and nitrogen metabolko I should be to strep the pouble importance of this observation place there ure ladation for a regulatory cyde at the phosphoglycerate level first niggested by Rapoport. Whether tbls hortono afects the phoepbo dycente cycle donn m Poul I remains to be los estigated. Recen itu observed in our labontory that crude muscle extracto cun conty ·464 Annals New York Academy of Saences 13 DPG4 23 DPGA AD ATP 3 PGA 2 PGA Fun 1 PEP Phospboglyconto cycle most quantitatively 3-pboxphoglycerito to 2 3-diphosphoglycerate * I am not certain that this is a physiologcal effect-perbapo its only on in 1170 artifact bowever we are sourching for alterions that may lead to the better understanding of hart fahre A block in the pyruva kinase (or molare) rauction, far oumple, oc umply low levels of inorganic phosphate and ADP could result in accumulation of 3-phoop boglycerito (3-PGA) Tho, in turn, would stimulate the convernon of 1 3-DPGA to 2 3-diphosphogłycerute (2 3-DPGA) Although the 2 3 DPGA would stimulate the phosphoglytente mutaso raction - thu would result in further accumulation of 3-PGA once the equilibrum o in frvor of 3-PGA formation and thereby further 2 3-DPGA matmulation Further mare, 3-PGA inhibits the dephoqphorylation of 2 3-DPGA to monopboxpbo glycentos" However once the block removal, the accumulated 23- DPGA would increase the activity of the 3-PGA mutex ed therefore, glycolyus. At the mmo time, inorganic phosphate and monophosphoglyter ato would be formed vu the 2 3-DPGA pbocphatuse Although we have demonstrated 2 3-DPGA accumulation in many times,* It is not known whether any or all these are under poch regulatory mechaniem. Perhaps the high concatnto of 23-DPGA in the human erythrocyte und its variations in dlake reflect such 1 mecbanien (2 3-DPGA may or may pot perform any other rolo in erythrocyte viability) It blikdy that the moed in eo effects thus for demonstrated with thyrouinc will be shown to have a comple result in the living animal, u anticipated for a long time by phymologat It must be remembered that most of our chomhal methode ue pot adequate to itveel mall but perbaps Important differences between roomal and abparmal ture uod that, to my knowledge, it to very difficult to vognhre crperimental heart fallure comparable to that in the human patient The carly Indrates the seed for i areful qmodatire stody of blocheucal panmeters in the buman pormal und abnormal myocardium. Port-mortem material may be useful in ach studies dance 1 number of components, per ticularly some ensyma, are relatively stable.

·Gnsoba Energy-Generating Systems 465 Referace 1 SonILAD, ET UT RA 1957 The proporbe a dear nbooockuoda prodood with cellular partide, ATP lit und quorparme a doamu J Add Chem. Soc. 1 3603 2 Lo, LFCC CAINI 1957 Brotheu alth cogan from undone dopbox phate glucor J_An Chem. Soc Torun J. BAR, 1 1917 The most breakin X TPC TOOL Z tamm, Hormon Fomathord 1(341) 301 #DLING, W&O Beoor 193. ( an hund.sockoud der Hof D (Adenoma 3pboqbarmoro) Biochem Z DS 110 S ONLINE, O 1951 A pocoonyme of wabouc lamentation. J Biol Chem. 10:11. 6 KURALLAX P&W FHN Vox mm 1963 Nodcoucke nchabor ID aduc ativity II Ranctoos n rok Cuculation Rouch, 1 12 7 Honou, B'FH104 Ideauerne tripbonphat broken dom dong tople mucketuitch Matore 174 1000 # TransTOY AT JANI RED I LH 4 Km 1954 Cootructe of punde without and done trpboxphat a crantide poophats Nature 1741 10.1 9 COLLO F ALICA, RFO &A TL Tax Cou 1958 Syotbadphopboerum by coroa frog bort and heletal pode Natur 17t: 1231 10 MODUIT, WFH 1951 Tbc tbation of poopbate bood hobokopal ter 7 Phospboro llenbolon 1 551 Jotun lopkins Preu Balomare, la 11. BOOTCA, EN 1937 Tbe dengan teraction of the beart to trophatun dung coperto hal thamuo defony. Bil Expu Bad Bled 11: 918 ngth translate troo, Consolants Barang, Inc New York, NY 12 LOND, ALIBLRA 1957 Ondavou reduction ato d nt krer oitochandele od the action of thyron Bochum et Bophy Acta 20. 613 13. Choola, 6 EN ITO 1957 A bacon on the telection of dutab I Diptabs . Thomas Springfield, 10 14 LIMAU F 1946 Acetyl phosphat Adruct in Entypolog : 231 Intar COO No Yak, NY 15 HALIT L 1957 The hydrolyn a 1 Jdpkop bogels ex eod by sci pompben. Plochum. et Bophy Acta si 134 16 COOLA, S&R O MAREAL 1954 Inryk decomposbou of the scttre loter Debat bu atrulha mathedo Bodea et Barbare Acte 14 440 8, DP WALLIC & I J GRADT 1933. Crtel trumfer It DDmahu od bacterial cor preparato Buochum at Bopy Acta 1T 150 11 GOLIA, S J CALATAC 'LB'I JOTTI 1958 Bbocfoon of Boopt) Actu Tbe 19 RAPPORT SE LUONDO 1931. Ghant 2,3 pop batam J Biol Chen 11663 20. GIDOI, S EA. E JOTCL 1958 Earyabeth and wolation of 2 3 diplom phodyconut J Biol Chem I pre 21. Romit, V W T C Toni ES GEDOUA 1957 The Worte propatia d you od mode pompboglycerx od on J Bal Chew DS 575 22 JOTC, B. K. 3 GOLLA 1954 Stwchos aconte 2 J duplopbator. J Brol. Chce 0 190 The placentation, babe 23. Tor, JCV Rooms Gul 1957 thatotthee of 13 hpboplagt com rod J Biol. Chem. I 17 GIDOIA pebberbed ·CONSERVATION OF ENERGY IN CARDIAC MUSCLE* Robert E. Obon and Daothy A Puteh Dupa of Drechstry and Crew Sand Rx Unity of Pesharh, Prork P For the past few year our laboratory has been engaged in the systemet cumination of the biochemial paths ays in the normal and fuling bart for evidence of a biochemical lesion that could be implicated in the pathogenes of ourdiac failure Since the abilty of the beart to perform work depends upon the intactacts of all of the blochemical reactions that lead from the oxidation of substrate to the final contracte proces, it xoped possible that internption of thus sequence ut any point might hand to cardiac falore Thb paper review the evidence that has been obtamed regarding the energetic of the myocardium m the dog uod in man under normal conditions and under two conditions that may lad to congestive hart failure namely thyrotaricods und kevere valvular durense. Bletabolxc procemos in heart muscle may be divided mto three general phases (1) energy liberation (2) energy conservation, cod (3) energy utlum tion There are presented dhagrammatically to IGOR 1 The phax d energy liberation 4 visualized us including those ractions by which the car bon-arbon und artoe bydrogen bond energy of obdurable whatrate da liberated us free energy In the phex cardiac fael which consists chrefly of free fatty Lads (NEFA) glucose lactate and pyruvate batracted from the coronary blood and 13 metabolized to a common intermediate, acetylcootybe A. The acetyl group condenses with onalacetate to yield atrate that has then degraded step to through the wellknown reactions of the Krebetrarboxylic ucd cycle to yreld 8 hydrogen atoms or electrons (H - H+ + ) bich represent the energy content of the wetyl fragment and art equivalent to the 4 hydrogen atoms unsocated with the acetyl group pho 4 that come from water added to cycle intermediates in effecting the ondation of the fngruent. The function of the glycolytec Lipolyik, und citric rod cycle miyma u thus to convert the bond energy of pubstrato annilable to the carduc muncle cell into energy-nch electrons for transport to oygen Thu phase of metgy lberttron merces with the next phase of energy conservation through the meche noms of oxidative phospborylation, some of which occur at the substrate land however the most important (quantitatively) of them occur m emociation with the electron how along the hydrogen trumport chain This depicted in VIOURI 1 u coupling the phosphorylation of adenocne dipbogphate (ADP) to the oddation of the cytochrome enzymes llith en 11mige P O ratio for arbohydrate or fatty unds of about 3 which a ell validated, the efficiency of convenon of the chemxal bood morgy of substrate to adenosine tnpbos phate (ATP) appar to mnge from 0 to 66 per cent Creatine phosphate The rod reported this paper wpported in part by mnts froo the mean Heart Association, la lorLNI the National Hart Insuitet (EI 100) Publc Hall Service, Bethode, did the L Imanscelledal Rearch Fud, New Bork, NI 466 ·Olson & Patne Energy Conservation 467 ORATION ICONS ON UTLL TIO + La M TOTO LOT C CAROUC FUEL CARDIAC WORK 4 UOC CU ADT ACT FOORE 1 Schep d merean arte mock (CP) Hora lo ardiac muscle ure relatively small (equrulent to 12 to 15 mg. per cent P) but provide sorte high energy phosphate buffer aapadty by way of the enzytoc trumpbophorylaxe The third phase of aardesc energets that of energy atiluation. The high-energy phosphate bond energy of ATP channeled int the contractile mechanlar and results in the performance of mechanical work Although there is considerable controverty about the model that best reprezenta tho contracte ryztem, the model presented m Fourt 1 bas the advantage of being dmple. In tha syutem, actin (fibroos) plus free you are postulated to combine in dlestole under the influence of ATP to yleld actomyosin und ADP Excitation of the membrane at the time of electnc syitole then stimulates (pamboty ya K+ shot) the energy rich and fully attended actomyod to decoctat into its component parts with shortening of the myofibriluod the production of mechanical work. The work efficiency of this pbase may be as high u 60 per cent, resulting in an ove-ul nod efficiency in beart muscle of 35 to 40 per cent. From the codeacet band somo yan spor me elaborted working by potball that postulated that coopestive beart fallure of the blgb-output type keen in uocatio with anemia, berlbert, and thyrotodicos, wus due to defect in myocardial cely productlon (indudmg the phases of energy liben tion and/o conservation) heraus coogestire bart futuro dl the lov-output type recoodary to valvular deux hypertes, Of coronary artoy docese mu due to deet in coery utilation, recondary to a losballty of the cootructile proteins to usimlate phosphate-bood energy or to uoclate and dis sociate property in the audac code Ith light of this hypothesis * shall tamude the data that bare been obtained in the mtetta thall emphade the events that occur dundg the phar o coerg Iberation Add cookerto in normal heart muscle and lo hart muscle from uploab and man with thyto torcode od lor outpat congestive heart falure.

·406 Annals New York Academy of Saences Ndebeline of lornal Cardiac Iwce Vuch evidence a avubble to indicate that aardix muscle at a variety of subenta o nources of energy including glucose lactate pyruvate fatty acids and to a leader calent acetate ketone bodia, and amino ads The client to wheb ech substrate coatnbute to the mergy requirement of tbe heart 18 ino u mftucaced by its concentration in uterul blood, umel by the rute of nutrition and the endocrine balance of the organion In add thon, once data obtained * titre on the ondative dupontion of pubstrates by cardiac muscle have been applied to the intact bart, it should be pointed out that the behavior of arduc muscle slice In turo dufan in keveral respects from that of the intact heart It has been shorn previously that the decay of the pyruvi orddex system in carduc muscle alles from nats and duckling sa loganthox funx ton of the time of incubation at 37°C Only at very short tone intervals after initution of the rubation of previously chilled wa could low values women hat comparable to those obtained in mtact mammalian hearts be obtained Additional omparative data on the etc Lion of pyruvate lactate ind glucose by ardiac muscle in litro and 1x inte are prescated in TIGURI 2 Cardu muscle suces from well-fed ducklogy were prepard u previously denunbed and incubated in phosphate maline medium with glucose lactate ind pyruvate seporeldy at vanous concentn tions from 10+ 120 mg per cent for 1 hour 10 an atmosphere of orygen The dependence of rate of extractxon upon arterial level that is, initial concentration in the madrum, us shown in the left hand plot For comparon with this, the extraction of these three substrata from artenal blood by intact dog LAODOT MW CITLID/-/au MW. OTOD //00 Mias fusore OT L LOVE un um Frar 2 Lunctwo of keratand mit by ardir mol or The kft baprl humo konto the chi vor al ardur mencie plic Incabut d lub tbex recte into at mors concentratom bombat In Der Wh u otro toybere for one hox The net band dunn dercnbe ibu bebanor of the mtact dog beurt al venous craintxone a tbon banta in astonul blood ·int 1 it - 1 Olson & Patnck Encia Conservatrın 109 myocardium us a function of arterial tration thor In th nghtHub plot The batter data y te dan II athet n ton of the lib posutroptnc and postprandal kan una rates ek ulated from the product of the arteron nam ne fx ah uhar te od tbe coronary blood for xured by th a tahrxu Certain dillenna In the mirin HTX hurt musle #tiro und i stw are imonedut lyptun tron of substrate begin at to concentration and fшга 1 dwutul arxcentration od glucose and lactate the s not nuk h tracted most npidly at krn concentrations at thett b d at threshold of utilimtion for each substante Ulthweh ilah ery but icro for pytavate (06 + 02 mg per cent) nu 25+01 mg per cent) the threshold be considerably higher for glue (5) 0 mg por cent) Abone the threshold the extraction nato rimer fun luss of their respecthe ar terlal concentrations in the range om vuntered in the poslabsorptly doy Itu poten orthy that these hoa * th the caption of that for pyruvate, bave Heeper slopes than tbose oben d untur maxle sket At Levels of blood glucose about 100 mg per cent the tracion curve for glucose become curvilmear approacting • maumur extraction of 25 mg/100 ga. muache/min, at blood sugar values greater than 250 mg per cent. The threshold phenomenon for atincuon of glucose and the 3-carboo ucida by Intact bennt 6 apparently lost in the dice Thut this torthold behav for of Intact myocardiumba function d un mtact cellular membna eh magrerted by (1) the loss of this behavior in tbe place, and by (2) mochfication of the throhold for sacose by changes m eodocrine balance in the bote animal. Unpar d i and Goodale a doha e thon that the threbold for the et tnction of docose s medly elevated In dabetes mellitus and restored tonard Dormal by hn treatment In pormal dogs treated with large dous of impulin, Hochd ha sbom p that the threshold for the conction of glacor an be lowered to 10+ 3 mg per cent Another important pour should be made regarding the metabolie bebe for la the fomer of louct heart muscle us opposed t cardlec muscle ances the crtnction ind oddanos o substrate ate nicely balanced m tbe ready cuate butter tbere si depanty between the amount of substant catncte add that oudired us shown in TOLE 1 In this cyperoncot aardbac moucke nice from bealthy dochog ere excubated in Kutuplicate lluborg tels with pytuvate bactate, and placok bogelet o concentrations ustang found in urtarlal blood One of these three pubsinto us taggedth C la orch pair of fuel, so that the production of C40, from a mbeirat The attaction of subcould be obseried to the protect of the other t Knate nu degured chemically and the udount of CO apxared to the center well over 1 20-poin period nu determined by eademoder counting of BC" Both the atraction and oddation of auch fubstrat art at prened in temos no Kuboa uced metaboliad pet my dry weight per bour It may be seen that whereas the total attraction of pytuat lactate Add glocome by tbe nices exceeded that obserred to lotact beart muscle th beleh ·470 Annals New York Academy of Saences Tu 1 EITUTO w Comoros of CAIXA DE ATE M CARAC Voter Is In Ap I Isto Oportet end 6 ) bande (catril ht perint Lithct Estructie 18 18 0 10. O 02 20 60 05 38 43 20 18 5 1 68 06 D5 OJ 14 Tyruyt Letat Chicou lon Percentar betrue a tructed andered Pertatap menu to ackcore about 100 10 96 30 cerit oodatlon of the extracted pubatnte to CMO, u martally reduced in wko Only Pyruvate us burned a napidly 13 til u ix it. Companons between the rotact beurt (postprandal) and the slkes indicated that whereus carbohydrate oncted by the nuct beart mus burned to com pletion, that extructed by the slka nos oridized to the extent of only 10 per The total rate of orygen consumption of the ulicas nos about so per cent of that of ibo intact heart, and the combustion of added aberate in the Hicas accounted for only 30 per cent of this tygen Courimption. It w be concluded from these crperiments that although » uTo Hudia o cardiac metabolom may partly reflect the qualitative aspects of carduc metabolism to there are marted duferences in the nato and patterns of metabolim u the two preparations Pyruvate a proxe substrate for cardiac maurcle both in nland 1 timp In FIGURE 3 ure prekated the resalt of 20 1* imo etperiment with pytuvate 2-C in the intct dog in which the apparent half life of blood pytunter 1.3 min Following the intravenous administration of 10 mg of tagged Pyruvate, the company sinus blood became napadly enriched with CPO, that remained hotter than the mired venous C, for nourly in hour The dotted line represents the cumulathe excretion of radioactivity to the breath that raced 30 per cent of the adonutered dose in 2 boun It boumated that about 20 per cent of the pyruvate odded in the first hour wu atabo Hted by the beet The wate of nutrition of the organism influences markedly the kind of rab- Arte wad for energy production by the bart Uoder phyddologal cood! tioru of feeding ud fasting the bart shifts frove the predominant otiliatlon d arbohydrate to the almost exclusive otisation of fatty acids us a source of cergy Under postprandbal condutions or after gtxcote Infoslong the myo ardium of both dog and man utilizes mainly glucok, lactate, and pyrovate us sources of energy and the respintory quotient of the heart approaches 1.0 After la o crnight fast, the pattern o metabolism shift to a greater ·Olson & Platnch Energy Conservation 1,1 IA COROUT US 9 CUFULINDRID PO ATIYTTY CHINA CHOR MODU OD PO centTED SAD vous 0 THE OUTS sop) The octabolum ol pynt 120kwut the Theobdres represent the charge la peaufic struy daartoe drone from corons RDD ud muru blood, respectively The dotted Wes the mult recono understrhy me breath to expressed (nett ortimal ) per od domated done dependence upon fatty acids," with a drop a myocardial repu tory quotleot to bout 0 R2. Gordon und Cherkesi bure tucently shoma that the poarce of ibe fatty acil utlled by the heart the Donostenbed fatty acid fraction (VEFA) bound to rerum albumin In their study of postaboorptive bumun rubjecu, it mu found that 25 to 70 per cent of the multloco oyen age of the beart could be accounted for by the uptake of noodsterified latty acide Tbone abuts are promoted to TABLE ? With prolonged fasting and with dubetes mellitus, the uptake of arbohydnate by the heart b further depreped with a decrease in tbe extraction coefficients for glucose pyruvate and lactate the myoordial respiratory quortent drops to on and the beart depeodt almost solely opon fatty acids for energy Danog transitions i nutritional stats from feeding to fustlof, the upuale of total arbobydrate and NETA varien reciprocally and correlatos with TARA 2 Met O Fimo Au FTTO Cror TOCARDIL Submit Umor DOO AON Credit 1 Y Huerta 0 per cat син Д С 1л NEFA All C 100 Pacenture 0, atumboe Kroosted for by CBH VETA 10 1 001 11 50 0 51 0 12 0 ·472 Aonals New York Academy of Sciences the obecned respiratory quotient for the heart suggesting that there are the major substratus for coer production in heart muxle Carbohydrate appears to be the preferred saborate of it u un lable and the animal in la" Hate therefore it appars thet arterial lot of carbohydrate the dative determinant of the fuel mittun tshen up by the beart and that the ХЕГ 4s Are чит." In thLamine deficiency in rats, dogs, and man, the myocardul ertraction coefficients for pyruvate and lactate are markedly deprened the respiratory quotient la reduced, and fatty and are utilized to Do incred ettent There also appuan to be reductxon in total myourdul oxygen coneinption under these conditions and a darordani relatxonahip between myocardu) otygen ettraction coefficient and coronary fox Thu Lilure to CLK OTTOTI consumption in proportiod to creased work load may be the critxul factor In the gener of the high-output congestive heart falore wen in med sitb berlbal Ndebalism of Heart Juscle * Dogs wik OTICASTS and Sam larda Disease Sinxe thyrotoxo (auke of high-output beart falure and val ular dieux a cause of low-output falure in min, it was thought of some interest to comput the metabolism of the heart under these two conditions in a mult able experimental animal The dog was home for these studies chiefly be caun of the large amount of data oo cardiac metabolism and hedrodynamics yailable in the literature for the animal uodet phymological conditions, and Kicaux boub thyrotoxxoor and congestive heart failure secondary to valve Lar doenhave been produced in this species In our study thyrotoko nas produced by feeding healthy domy 1000 mg he day of ISP thyroid porda for two to four months lleight loud in the presence of martedly focused caloric intake markedly in Trased total oxygen consumption mild tachycardie, increased evitabilty increased metal temperature increased hematocnt and incraiud levels and turnover nta of protein-bound rodine ae noted Chroom lov-output congative heart fallure characterized by edeme, axdites, wealder, reduced erecte tolerance ardlo- and hepatomegaly and elena tion of both nght and left ventrcular end diastolx filling premeures wu produced in dogs by surgical vulpon of the tncuspid salvo (TI) and steadis of the pulmonary artery (PS). Although initilly mght-side bart Callure induced in the way eventually becomo genealued u indicated by claration of filling prettures in the left ventncle and a positive inotropic re ponse of both hambers to digitalise. In our 4 b hon i kna o pro mre inang obtained by candoc atbetenation in two chauben (nght xuncle add nght venincle) od of the hart and three grat vesels (coronary doo, pulmonary artery and lemon) utery) of a typical dog before nirgery uter Induction of ladure uod after treatment of the failure with acetyl stro phantbidin, 1 rapidly acting digitalb drug* Right ventricular eoddastola pretres, markedly devted after the onset of fallare more reduced within ·Olson & Patnel Fnergy Conservation 473 CONTROL FAILURE (TI/M) FAILURL (TW) BEFORE DIO IHLAFTO 2 cu u DOT ma Hd RA wu -10 10 TV 30 bmw -10 O VO -10 F10 0 MA 180 wwwwwww LIO -60 TL + Reprezentativo preverre trem tryght unck, corona Hatice, plorary utay demoral artey dog beort er ofte oport butas, adufta vartent of the boat whiro th t at Darts of acetyl bar coods by Fuad trung ·474 Adnals New York Academy of Sciences one pour ulter admiration of 2 at units of acetyl stropbantbrdio with 20 increak In nght ventrcular systolic prepare and an increase in arduc out put. The ardue metabolism of a kta of thyrotor dogs and of a mno o doga un congestire beart fallure due to shular diseax nu compared with that for a verlas of bealthy control animals Cardiac catheteration with intuba non of the pulmonary artery nod the coromry rinus 18 performed on cuch animal modo Nanbutal-del-morphine anathala. Premura in the major veures and the chambers of the mght heart were measured electronically and arduc output u Dergured by the Fkk principle Coronary for mus detomined by the Dutrous oude stuntion method Artonovedou duffer ences of otygen, carbon donde, glucose lactats, pgravate, and nonchenfied fatty acids were meared manometrcally or chemically on sample of arterial and coronary gots blood After audac atheteriration artificial respuntod mu mitlated the chest nu opened, and the inimal sacliced by creator of the batting beart Determinations of total add-soluble płodboras, truc inorganic phospbons, and adenosine triphosphate were made ou quch-frozen Mmples of ventricular aper by i modification of the barium fnctionation technique proposed by Lepage and Umbrelt Values for renting cardiac outpat, coronary blood for curdluc orygen uroge crprewed us ml min 100 gm and ac, for Dordal and hyperthyrold dog, 1od for dogs with congestive beurt falure scoodary to TT/TS are presented In TABLE 3 It may be seen that whereas the cardbac output s decreured in chagestive heart failure umoclated with valvular diseas, its us madedly Bugmented in cxperimental thytotodiods in the dog es in man* Coronary blood flow and Dryocardial orygen mage were unchanged from normal in congestive heart fulure due to galvular disease in the dog, but dgnificantly Augmented in thyrotorkcon Altboogta Bingo reported Do McTex to the Orygn concompton of the myocardium in man with hyperthyroidion, con- VIDcing evidence that coronary flow and total myocardial ouren consumption are crand in human thyrotonicods bas been presented by Rome and hos CO-xorters Data on myocardal substrate and otygen attraction by postaboorptive normal od hyperthyrold dogs and dogs in congestive heart failure duet TI/PS are precated in TABLE 4 It may be men that the individual arterial Tereta and arteriovenous difference for glucose lactate, and VERA are not Tan 3 CALDAC OUTFT Cour Blood rom, AKD CARDUC Onion UWE MY Yoru DMM Thon Doos A n Door CosODSTITI ur Fawr (wy ortal Curt X Cybo od 1 Caymay pet in d IM 7 Vonn CHIF (TI/15) Hyperthyroll DO 202 + 12 91 +9 1 + 10 + L + 01 +57 121 + 6 12 0 + 0 7 10 9 + 1 227 + 3 10 5 + 0 10 $ + 20 25 + 3 14 8 + 16 17 9 +1 +13 + J ·MONOGRAPEIC, PUBLICATIONS or THE NEW YORK ACADEMY OF SCIENCES (Litty or \ATURAL HISTORY 1817 18 ) (1) Tho ARN I focta o series) established in 1823 contain the centific Cootributions and reports of researches, together with the secondo meettogo of the Academy The artido that comprise ach polube er printed op rately each in its own comer and are distributed Immedutely upon pablication, The prices of the ropante eroido depend upon their length and tho mlimber of llastrations, and may be ascertained upon applikation to the Executive Director of the Academy Carrent Damber of the AVTALS ure kent free to all Members of tho Academy deuldog tbem. (2) The SPIGAL POBLICATIOS established to 1939 are spoed at inregular interval u clothbound volumes. The price of auch volume will be idrerttacd معنا ه م ء (3) The Alps.coms (quarto wria) established in 1895 are issued at locular interval. It is intended that each volume fall be devoted to monographs relating to some particular department of ence, Volamo I Part 11 devoted to Astronomical Merdar, Tohume II to Zoological export. Vo more parts of the Mermoirs have been publi-bed to date. The price is one dollar per part (4) The SITO SCACY OF PORTO RIC AND TE VIRGIN ISLANDS (octavo rena) establubed In 1919 ghas the detailed reports of the anthropologlar!, botanlal, geological, paleoatologlal, bological, and metoorolograal Wirey of these and Sub captions and inquins concerning curent and back numbers of any of the pabllation of the Academy obould be addrewed to EXLCUTIVE DIRECTOR Than Tak Acgiaty od Samu 2 Ead Suhted Sirro Yelort 21 AY ·474 Annals New York Academy of Saences one bour after adminlgration of 2 at units of acetyl crophanthidin with in incrax to right ventncular systolic preure and an increase in cardiac cat Put The arduc metabolism of a remes of thyrotoxx dogs and a varies of dog in congestire beurt fullore due to valvular dikus mu compared Ith that for a wrics of balthy conud! animals. Cardiac atheteriation with tube tlon of the polmonary artery and the coronary gou wu performal an auch animal under Neubutil-dal-morphine anesthau Prepud ta the major Veels and the chambers of the right heart kere masured electroplus and cardac output sa masured by the Frok procłple Caromry low determined by the nitrous orde atunton method Arteroronow difier mca of oxygen, carbon dionde, glucose, lactate, pyturite, and donestated fatty acids were measured manometrcally or chemically on amples of arterial and coronary cous blood Alter cardiac acheterization urtificial respiration wu Initiated the chest mes opcoed, and the animal sacrificed by exchaon of the batiak hart Determinations of total acid soluble pbosphorus, truc Inorganic phosphorus, and adenosine triphosphate were made on quick froxco Amples of ventrkular apet by a modifiation of the barrum fractionation technique proposed by Lepage and Umbrelt Valdo for resting cardbac output, coronary blood flow cardiac oxygen suge capretad u ml min / 100 gun and uslo for normal and hyperthyroid dogs und for dogs with congestive heart falure acondary to TT/P's are presented in TABLE 3 It may be seen that, wheras the arduc output a decreased in congestive heart failure owoclated with valvular dureas, its ne markedly Augmented in experimental thyrotor con's in the dog u un man # Coronary blood floor and myocardial otygen toege were unchanged from portal in confentive hart failure dur to vzlvular duase in the dog, but agnificantly augmented in thyrotocoas Although Blogreported po increase in the orygen consumption of the myocardium in man with hyperthyrandum convioane evidence that coronary for and total myocurdal oxygen congumption Art Deus in human thyrotoxxcool has been presented by Rowe and bus Co-vorken Data on myocardial substrate and orggen extraction by postabsorptive Dorula) and hyperthyroid dop and dogs in congestive beurt future due to TI/TS are presented in TABLE4 It may be men that the indindan arterial leves and arteriovenous difereticos for glucose, lactate, and NEFA are not TAJL 3 CALDLAC OUTPUT Concu BLOOO Flor AD CALDUS Ono UOL JY NORMAL AD HEITI Top Dool A n Doon CoopTITY HEART FALL Mob Cart CAY wa ch Norm CHI (TIPO Hyperthyroid 2012 + 12 91 E9 3 10 98 + 15 4 01 + 37 121 + 6 120 +07 10 9 + 1 257 + 10 S OS 10 5 + 7025 14 8 16 17 9 E 1 CU ·Olson & Patch Energy Conservation 477 tahalır diyak no changes in (P OP ATT kesels mete Doted in the qoutaneous wimy myocardiam of the bun lung propantio. Since CP leveto fall la raponse to allore di energ liberation (anodia) or teto conservaton (D\r) in the beart-lung propanzioni probabts that no blochemical datur bence of tho type criate in ather f ilun due lo valvular doar ar thyrotocols. On the other band these datud pol ghe us an loht to the mechanien by *hurch tba almente do use ongull e heart favure The shift in orld.the metabolom tomard tbe utilan farty acids in thyrotodone that also occurs in diabetes, by itself hardly would constitute a fufficient reason for the myocardren to fail Turther study of the problems in progresa. As regards the blochemkal lesson in the myocardium of dog with cong the beart failure due to valvular down videixe recently obtain d in our labora tory Rures thet a bia hemkal df i my ost in the contractile mechanbum itsell.* * Vyvan rus bolased from the bearts of animals in lalore and Altable controle, ngorduals punued and physkochemkal characteralba eccomphshed by Hudy of ediment son onstanta, difunon constanus, partie pecllir volunci, minoso Oks, nd hight-rattering behavior of the two preparations From there onstunts th moleculer raight of ardiac myosin from portal dog de alunal dt be about 225 000 berou that from dagen bo faulust due to alvular daraur us found t be of the order 750,000 The dou are content with th xu that a abnormal Hable pregate of myonda Al formed in aarduc future od that the prevents the forma ton of an tomyndon with DOTTOal ontratie property There finding art coluistent ith the oburstions that actoryou prepunica from ſaling betart muscle of dogs und man shor certa boormal properties and reduced contra ulity Sonery Cardac energetics may be divided int three phant (1) erg lubention, Under normal condition (2) energy conamaika, dod (1) sieru atuhiation mechanku wart these cycles mehr Oont chemard encry deboency us high us to per ent Glucox, Lactate, pytunate and donaten- Apply th ball of the corps for cardiac muache, od the state of muthion or endocrine balance may shift the aubetrete mixture at ted by the myocarduum from preponderantly carbohydnate to preponderantly it Proloogd fuung untreated dubetes mellitus and thymtorkou lood to zhut th pattern of myocardul onderson to a greater utliation of fatty acades lo congestive bean lukore is dogs and men there is no eguficiat de er ihop 12 cultive metabolism A tudy of the darthibution of poopphoru amor the gency photo et compounds has reculed angular back of contun ht it parture frono pormal o both thy rot w dog and do falurt The dat great that no stoor buxhetra) kuna phax of coergol beration of om atmo in other thyrox Qou or Although th call mal dit quest that an abnormal ty of anine moun ant anonals to lulure due to rubar dekat ro clear plancon of th chile terwus effect of thyrot teosts upon myocudol fimction pparent The trata bed fatty the مه dmc ماله | kom wtput congestne han i Jure Kcondary to ·478 Aonals New York Aademy of Saences cffects of bypermetabolism ind hyperthyrosdisa upon the similation of phosphate-bond Cetty by the contactile proteins and the effects of the con tncule protams themselves should be further studeed Ackurlausais We are lodebted to Virginia Bartlebeugt, Samuel Barfield, Dorotby Territl, und Nullam Steroski for dlful technical trounce References 1 LADY HA 1952 The Biology of thosphorus chips Stat College from East Laming Nich 1 TT-GTUDOTTA 1917 Chentary of Under Contraction Aanderopc Pro New York, NY 3 Stry-Toron, 1956 Socce 114 873 4 MOLALTI, JIFBOTT J J Blon & T L Hu 1935 w Ren 161 475 5 HADOR, J&H E Houn 1933 N ture 175.00 6 OLAN REUBS m 1931 Blechase 1 2 7 Bono, RJ 1953 Metabolvan a tho bumun baut w Jut Smu Hoquial 0-100 8 MILLAL, O NERE OLSON 1952 J Biol Cibem 129 4:57 9 OLLOY RE O H Plan, ON JULLE & F S START 1913 J Biol Coco 1TM 459 10 COODALL TT&D B H 1953 Circolaton Rovarck 1 502 11 GOODULI, W TRE OLIN & D B HASIL An J Med La prt 12 Vroun, I M G A STOEL, J NDLAD & R B20 1955 An J Xad 13 GOODALL TRC OLOT #DB HMIT 1951 J Chn Imart 30 60 14 HACTII, DB Privat cumonabon 15 OLOR, REA RAUSVADB SITWANT 1957 Fedontoo I'roc 16 219 16 COCOALE, TDB HIL 1953 Lotha Roarch 6 309 17 Brwa, RJ A SMAL, I UW EI OO 1954 Am J Med 16 501 18 (ordoy, RS * A Cim 1956 J C... Iment 36200 19 HACIEL D B T GOODALL & Kimon 1953 Am Heart J 4 3 20 OLO, RT 1058 Federatico Proc (Sappl No 2) 11 21 21 Duo AT SF BAHIA 1916 Endocrootao X. 20 BAROK, ACBB Rot EGS RICKARDON 1982 Am J I byl 11 4 23 PATIEK DIURE OUDY 1956 Federation Proc 15. 115 24 OLYOK, R E G Rots & SI IL LIAMO 1955 Circulation 14 755 25 LITO, O E&U UM 1947 Metronet ne Techoes Boren Vonne spolu, mn 24 FouTTO, CAIGA Fano 1930 Bull Jobs Hopkins Hopp 6 203 27 BNG, RJ 1931 BoD NI Aad Ved 5 +07 2 KON L GGHHHOLTTON A BROTTY HT TAX J F B &CW COTTOM 19:56 J Cm Leront 15 271 29 GOODAL, V TRE OLOYD B HATA 1950 Tedentom Proc h 49 30 BOTTL, RJ PLANO PF Oroop T L O 1956 Corcohtron 13. 1 31 LAXDY UIA&GFALD 1955 J Biol Choo JOU $35 32 LAZDY HAKO MALTY 13 Coord, CALLINGD 1957 J Bal Chara 3 561 54 F WALG E H IB TUTUN 1957 Bnt J Pharmacol & Chemotherapy 12. 270 35 TAWAL G&B TOTT VI 1957 Bnt J Pherocol & Charcothpy 1h 773 K ournon A 1947 As j Ib 110- 733 37 O REI Furry I STD V 31 LLO 1956 Ja Ime Hh in 38 STX HE E MUDRC OUDY 1956 Tederation Truc 15 (1) 10 Dry FS 1959 Cuolation Board : 221 10 KLO KR JHMO 1958 I Cho I 7 +65 ·Even Olson & Palck Energy Conservation +79 Dixos E EUL BOGEN (L mirosity of Puberg Prasowat Pe) Bly remarks concern the characteriatlon of mayonn alted from normal dot beans and be limited to the pomi thu h th no have been often neglected in phgual crants First, the heart must be pomal bell phonological criteria, such 4 normal times to thi, Dormal und kontamod normal or The cost of the olmalmost be opened under standerd surpal techniques, respiration must bemutated, and evention must be napad ind to officient that pormal batt comme or the crooked beart has been dropped to ice cold water I short terminal venincular fibra Liiton all prevent the folation of pommal adec myosin Second one must for citrumme pains to parlty the botaled mry out so-called cry line mo anart contammated with wome Kocmyusin, and a pretpon of pnpur in ultracentrifugation and repreplations as Decorary for the prpuration of mos that a bourogenicons in al respects. As a consequence the yacide of the protete ure rachat mall, of the order of 300 to 400 mg 100 go of tre Finally all physicochemical Daugirements must be carried out to Ty delat solutioce, in order to make cettate that entra polstrous to reto protetu oncentratua ere paningful Fathere to do so I result in a set of pbrerochemical contents that produce erodedus molecular aghu add shape constants tuh these quenza in mund che chanctoured ardiac ragoun from Dormal dog beurts by Kolumentation docity sedimentation quilibrioma Approach to redumentation cquurbrauen, nacouty partal patetic rolume, free diffonon, sedimentation borundan preeding and light mattering mer anto of 6 12 SD, of 2 +6 X 104 Uwing our values for h CA. Acc. and vol 131 Doberdar eight of 226,300 s akabatad From medhentation equilibrium and approach to redwantation equilibrio per meats a molecular weight of 223,800 $ 3000 obtained from the bene datt, unell us from the lowme poety number (abo called mtrtek ve cout) of 50 CE! with the shape of the molecule can be descobed us a pro bate cliproud mob on mul nato al 24 1 and frictional rato, Ile of 2.14 Uning the retury blue line, the refracto under facernent of mayo 0 206 and the light cuttering molecubs aeght = 770 000, sbuch a greca nel with the other maritet U of these mattirements are conabteitth. model for under mayoon of coul 690 X long and 28 A thick As Olson bou voderated, it vu found that these pbywu constant are mark odly altered for my ORLD Dolated from dangt conyentne heart lore Theme denn ill be presenta un fortbroelog communicato As a detence suodant, rabbit skeletal mondo e porpered me the same way and studied Moterohr to contains of companble magnitude menght cakulated from redwordt toe and difusca data and from that natteny date for rabbit scketal sapoan ere bo upreement I bout 150.000 ·ELECTRO TRA STORT IN CARDIAC TISSUE Thomas P Singut Edd Biwaſ- Vakel Retard de la rue Dart Wikt Introduction The normal contractility of uria muscles dependent on a complex menus of events beginning *ith the entry of ordiable substrate into the cell and ending with the reverubte Inndommation of actomyusin An niatupun ar major alteration of the procen anywhere along its path may be espected to result in important changes in the efficiency of the sort of ihe beart. Since tho aries of interdependent and mterdigitaling brochemku events sob viously enormously complet, for the wake of omplicity on dokusang congestat heart docar we may arbitranly divide it into 5 groups of racibors (ACHILL, 1) (1) The end of erymatk reaction whereby metabolites are converted into CO and HO in other words, the chemicu tnnformations of the pubunte Whether we ure CODcerned with the burning of carbobydnte or of fat the mafonty of the corymate steps terve merely to rabape the chemical structure of the metabolite in preparation for the orative step during which energy will be liberated It a logul to eumune the ondative steps in metabolam m more detail not only became they are the ultimately useful steps but beause they are of a much greater order of complexaty than the sonondative stop preceding and following them (2) During ach oxblative ucp machinery must be provided to trap the cheraxcal energy liberated dunng the oddation and to per it along with minimum of low to another entymatic syrem apable of genenung ATP I The machinety performing this lash more or less similar in all the individual ouldative steps of metabolism, and it a commonly rdared to the electron inansport system In an organized metabolic synem it must make functional connection with the dehydrogenases uskociated with the abstrate on one hand, add it must be mumately linked to the ATP generating them on the other hand (3) The ATT'-generating or oxidati c-phosphorylation system out then trap the coegy libented during dectron transport and uulize it for the tyttherls of ATP ATP then, may be directly otilized as the primary fuel for muscular contraction and for synthetx procentes, or it may be converted to other comopounds in equibrium with it (cratine pbosphate add vinod nucleo ade inphosphato) for purposes of gonge or for utdation in energy-requiring tranformations The orh dexibel na to parer supported by ts from the ton lart Inett Ne llomb Stryke in the prendan Ilmut A means bet to the oth oN Rreth, Department of the day wil the IB Forell da 1 for 1 dal Rant, bar 1656100) Internet de bedoeling is one to lose weigen helt andre les manteneSDP atramane ireplace ITT Bird Loukahl BIL Centine Idqboqbopaneline uchronik DP reducel DI DEVI y tochrome oth n we daneduk, I? borba TIIV redoved IJIN TANÍh tnphopbopy the recolle TPY rohre TPV TTCH contract 53 ·Singer Electron Transoort in Carduc Tissuc 181 mu i (1) 10 nunta con o metalokta th otisalt oodation Artis och) (2) I met nuk Indung (ket (3) Cound ! (oulain baton) (4) Thứ (sctom On UN (3 Jonur oth (*) The actoryoan-AIT wm then onverts the chamaal court of the terminal photpuste bond of irr into mechankal work (5) The latter process App i bx oot olled by changes in the corregin tions of duffwlole ons during the craciu proces From tbc viewpoint of th banhem! ny one of thek Sproccio may be potential wes of a metaboix dmurtunk that uumtely aspeenar itsel Ai claxcally recogniable onnan lain 11e have omitted from thu hat a number of other futon thu o usly infuence th mechanical work of the heart woce premiling op mon does not regard then u common Cup in the etiology or ibe daeus Thus, although on Apply ruy be charty Imaling factor in the entire sertts of ordative events, pertkularly once cardiac sacle toleratos only about one bith u mx Ds debt es doch skeletal moecke, the nute of Os utilasinua the (xuling heart may be normal or even higher than ba pormal hout Alterions in the permaabihty of cudar ella mortabo btes will not be opened ber unce Bmgh Audies of interual-vedoos differenca m coronary verkets show that the entrACILOD of a important morb Antes from the soul within Dormal Limiu in myocardul mare Similarly lul te of nervous corral dors not seem to be regarded a primary cause of congen blure although experimenu on denervated beart (ench heart-tung prepar cons) pount the importance of terwow control m determining the car yol buoni baxt Among the processes ved n&HEVE 1 the third lourth and fiftbare de Costed hewhere in th porcorupt The tru top the maymak tepu in the aubolam of buttato purtxuberly of arbobydratas, ha theo been die ccd Horon sinxellt remember that arbohydrate metabolum ss pot The oridation of th aly traporlent source of energy of mammalun heart flacone ually (ted to be of m nor inportance in nopplyve ibe Dergy Io la fatty od uod lactate ondation requirments for ATT monition are the primary source of energy for anda usare and arduc glycogen Day bo tegarded o emergency foel TABLI! taken from the root of Bing shown that only bout 15 per cent I the opel of the normal humo beurt ret t th uchamn of carbohidrat und Handrute mtermedute detailed decoroon of the peihnay of fatty ant Sput does not permit settudation that has ber defined as a call of the outstanding investigation by I) (ree I'm malty o limonun hladison Ilm Shos Tea Jork nd F LYBET II Planch Im ut 1 Zelke heme Su Khe Germany Honey once wity and oodattoo sa major source of extra for ATT FEDTILon in beart touch Un erat collere of Liedk oe New lod i ·482 Annals New York Academy of Sciences Tanr 1 RELATITT CorpoTOYO CALIDAD NOXCALOOTDATE TO TOTAL N OCULOLAL O, URLOT Curbek cocartel deste 17 90 054 Glucose Pyrorat Jactat Toto Fatty acids Amudo roda Ketona 16 46 67 0 5 6 43 769 341 90 Rqroduced by perman of the I B Seunder Cocopany mention must be made of the site along the fat catabolla pathway where energy gented for ATP syntheos Fatty acids are ordized in the form of their coa derivative in a series of reps leading to the formation of a fatty acyl Cal therlar with 2 carbon atoms ken than the parent compound und a mole of acetyl Cod RCH,CH,COSCOA - RCOSCOA + CHCOSCOA of the enzymat steps involved in ach of these Boxidation Hep, 2 ure oudative m mature and are probably coupled to ATP-forming system. The acetyt Cod generated m fatty and aubolism is then oudured yta the Kreba tricarboxylic acid cycle us the acetyl Cor generated from arbohydrate oddation Tho atabolism of ketone bodies, i wpplementary source of energy for heart muscle, aloo leads directly to the formation of acetyl Com Thus, hule the terminal oodative stopa in carbohydrate and fat metabolum are common, from the standpoint of energetxe, fatty acid and carbohydrate ondation differ in one important respect, namely that in fat orldation a relatively larger fnction of the entry 3 released prior to reching the stage of the tricarboxylic acid cycle This brings us to the second topk, the electron transport system, the main subject of the paper What follow is inteoded primanly for the nonspedalnes m the field and, therefore I have attempted to keep the discusion on relatndy ruople level, minimizing the presentation of onginal datu at the expenue of offering very lutlo information with which most crusyidologbu are not already acquinta Fledrux Trorsport Wechseln During the acrobte atabolbam of wory oddiable foodstuff at each oddative Hep a pair of hydrogemus apd of electrons uro removed from the metabolite The passage of these clectrons and bydrogens to Oy throaghu seria o en the electron transport cham, entails the stepwise liberation of cay (BCZYT 2) Each portion of energy liberated us in going from Y to Y t linked to the ploophorylation of ADP t form ATP In aardic taste the entire proc ortun in the Arconant. For the purpose of this discussoo xe may cbally the important mechanisons of electron transport in heart under four major headiop (airul 3) de learn more about these themes, it becomes incradndy apparent that ·Singer Electron Transport sa Cardac Tissue 483 1 Heulont Jetabolile B Jotaloutt Al Dicalon Urun I 3 An I c [24ORT (1) proton-F aproces - Cylorbros (2) Debt droprotocroot en -O 3) Dehydrerbree-DP-( torbrorento 1) Debt dructur-TP - DP - C fortsatt er the umilarities among them are mach more compreson than at the differences. The first type of reaction representa th oadation of a ribalrate by a favo protein, which then requestbe medation of a rtond fiberoprotein, the therton transfertlog favoprolein (ETF) lunch i to the cytochrome synem. The wrond trpe include ytochrome reduring dehydropean that on trailer favoproteins I melahile oprocesos and are apparently crpable of pertung clectrons port or less directly out the cytochrome them without the mediation of a distorable COCOTTA The told type of sitem urus *th the reducidon of DP by one of the DP requiring d by drogenares the DPWH may then be dowonted and poned on to cytochrome-reducing deb drogenske system of the second typo that accomplomban is noodation Io nome matascos tbe utaution may be more compter the reduced DPX may be first reqared to react with bound marcosomal Drpnor to poming the cytochrome pathuty The fourth group louloda the TTV-reducmg dehydrogens TPVH par be reoudied in BANDET analogout that of DPH but there considerabile cridence that in beurt in order to Lak the reodude of TPV71 to be forme to a ATP o altogetbet dulcro path y open to that mohu u tom hydrogenation bet e DP and TPVH the DPVII thus formed renerale the energy-noch pboxplat 1 shal Dot come each of these pathway in deta) Ondas ons The mutual step in the dehydrogenation of later al Cat the won the bumet maled ampunx's in Amst time In sch41 ) (lyze ty mua bomolques to utythes it Bixbd der Anwag ibang conty in mind I spenny for the buin korth fthx tetty urate Hibouch the mrekular enzyty under dixwoon ha been bouted from laser thert uitke doubt that i unitar kuation tots n beurt La lact, beurt mitochondrua hne been a preferred source for ·484 Annals New York Academy of Sciences SO4 PANY Or L.LECTEON TOTT V T UTH DTOUDUTTOM O Tam AL DETTATIVTS OF Cat (c-c G rity Colcrc 1 - ETT Acceptors cra Y Iron Barnett und Crane reproduced by permion of The Jobins Hopkuns Prem the solution of many of the enzymes involved in the oodation d fatty a hoda The peculiar property of the 3 debydrogenates (G I and Y') u that they are unable to react with electro accepton at ipprecuble nata in the abecoce of another flavoenzyme ETT wbore fuoction to medate dectron transfer from any of these dehydrogenases to electron accepton The natural electron acceptor of ETF is not koond but in highly purified propenations, reduced ETF readily oddled by artl cual accepłons such a dophenol dyes, phienadne met boriliate, and fenkynde more som ly and to a vumble extent by cytochrome e ind only extremely slowly by 0 In this lost cytochrome is the only effective detonal electroo tnnder agent but it is doubtful that it accepts cheirons directly from reduced ETF in intact rysterne One of the rusons for this that the interaction of reduced ETF with cytochroexe c is very omplet (KITUT 5) Beinert and Cure who fint deknbed the co- The have found that bưe hugbly punied preparations of reduced ETT Trapidly with dyos and copoderably more slowly with ytochrome parations acquire a high stivity_toward cytochrome c after acid so treatment and more further fructionation, but their activity to wie dyes romana ether unchanged or dectina To muke the atuation or more comples, the same punfied preparations of LTF prior to add (NEW,SO treatment, contain a high daphorns (DPVII anul TT'AH dehydrogenus) actty lopend dyos but very little ability to SIDES TOT AC ITD SOITATI RINDTIATION OF TI VOTTAL IXDDNLT O LTZ # VE TO TOOM AND DEVI wm TPNH DOLIN CLECTION T DU ROINT OF T I DNUNC HD TREATYTA Cytochrane G) TT Dum ITVI DPYII Edort kol trainett 1 Ludophenol tochtone 3 ITT D por vieradd todoptenel The ) Banert u ·Singet Electron Tranqurt in Candiac Tissue 195 be the onldatron of redord pynidiot nu leotides to tochroase Alter xod (VHS treatment fractions but be obtained with bigh ETF and bu teption &tivity and other in *th high daphorase and love ETF Kihlly the latter ut aloitt fox hrode rdtk tara la othus Nords, (\II ) ) treatment gamit itre puntlow of LTI and DP\II matang acth likes to a large ettent and the ski the ability of both types of De to react with cytochrom Vi u or the tno prepanthous re tabiano conditions wmilar to those that nted prior to und (VH),SO treatment that k, inhibition of tahtomt redu tion by ather ET171, or by reduced pstid we meteryxde 11 m that is the original animated entryme tbe oodation of DPH and of Latt und LETI by ytochrane u mnotu ally inhibitor On further imergulhan the might pronet be wo type of control mechaphm that may ho orker ble importa in (bsues where BY mention h largely dependent on fatty and metabolism The branding dendrogen For many yan group o oddu bag enzyma knons to pass eher troms 1 The tax brome fyern without the beauft of medot too by mobil soluble co-conto ba e been referred to u Splochrome redung dehydrogenases lo hart the mou mportant nem bers of the group are the enviou responsable for th ondation d Accurate and a DPH Because of the key position of the former utb Krebs cycle end of the latter i ompling og ridine-ouchotade detty drogenases 10 (), ty Sure the majanty of meng generation w ardoc idove unohto the participar tion of these imo enxyme syntes In fact in all clubood e art Dot darling maht o enzy toe syuems, but with one #tructural and functional unui inters laking some bere deur the level of vtochroated whicho called the electron transport chatn (SCHEME 6) lammalan heart bas as yo been the famous material for the Etudy of the chals of carytach, and much of bat we loon huo beto banned from partments with beurt preparations In heast Arco ma and even sub postochondital unu derved from time, the electron transport chain behaven In 4 bybly organized ration of Corpo rith definit kracture luct, for many youn it ius bebered that trutura organlarimon pays auch toportant role in the utmer of the wet that porficavon und sobation diu mdidual cnrymo mould be impoble Although, und a few yean th andarm fuu 460, attempts to solat tbe opocent of the system met are much nu lented about the properties of the whole from pectroscop Hudla uod from kidere esperiment on the ordata of criate and DPH The components and no he e been recognised on the best of their Back n TUT OF TO ETA TXmer Gau Lo de prolan SD) Cytt Crt Cyt 4 - 0 DPH - Iron Maphoto (DPAD) ·486 Annals Ney York Academy of Sciences specta in pcb particulate preparation. In addition to the hemoproteios Jated the stence of a factor actiog between b and has been topatedly proposed on the bem of the inhibition of the chain by BAL, antimycin," and certain repéthoquinones, xbxh te thought to combine with it bonner direct proof of the costence of the factor has not yet been found. llic shall now briefly survey the patient properties of this chain starting with the prury dehydrogenase Succinic dehydrogenax, wolated from heart us a soluble protein, contains one molo of farm and four atoms of Fo u prosthetic groupa" The flavia moty (FAD) s unique among flavopro- Teing in that it is jouted to the protein m such strong Inhages that it cannot be mepunted from the bulk of the protein except by protcolyes of the batter even then it does not come of FAD The participation of the flavi in the catalyos umambiguous its the primary oodant of excelnate and the reductant of famante The four Donhemin Fes are more lonely attached Kloce they are xt free upon denaturation of the protein Their role in the atalytac actrvity u will uncertain Atlas ID of the four Fo arms are cuentkal for the activity of the woluble entyme, bat despite extensive invest gation no evidence has been found to Ripport the power bulity that they undergo ondoreduction The atalytic properties of the dehydrogenaxe are quantitatively the amo in the solated highly punfied rate us in Marcosomal preparations the enzytac inhibited by malonate and other pubstnte amlogues but not by antimycin or BAL, it actun in a forta of low catalytk activity and may be a trated many time by contact with Riccmate or with analogues of the pubetrate u In one respect the soluble cryme bebavas very differently from fu counter part in particulate beart preparations upon solubilia bon the reactivity toward many clectron arrier i lor altogether and it is grantly reduced toward others Tha ba largely dat to the reparation and removal of other factoo nta (that , other protons) by the solublimation procedure The relative reaction nito of the dehydrogenase 10 z tymaal pertkulaie propuntion of the organized respiratory chan from heart are shown In TABLE 2 Phena de methosulfate maare the full actmty oodation via the dectron trampon chain, popple Dented by anternally added cytochrome c meriros most of the activity dye like methylene blue barure only a fnction of the activity and that only TAB 2 ACTITUD Or Kurfuru PDALAITOY 100 TAMILY ACTION AT STC (V VALID) Portu d Krtiny 100 3 Produse petbontato 0 O (with added cytochroon Faths olde llett ledo bere Boot croyl bloc Hydroom 5052 2J 52 : 22 13 33 23 69 Pamethasohat takes us 100 ·Singer Electron Transport in Carduc Tissue 457 T! Loum RATON Rumi ox Bfrt luut You DorprooIVI AT 25 C. Lite kterier Laatu DOO ! aww, os. 007 Porus oke Cockror 001 i pro 2,6 Ductelorophenoladole 06 hoc pothuate 100 2 (9) Tak ctbook 1 ion that can I blue 01 Vallcarlos S Tobe 2 Hydroty upbtborrone + bot bare for - Sa ! FAD ++ pot naland Du befrute 11 DP 01 (qprea) Dot poured cochologen Theatre-crboss lite 0 A W DIA are compared th the forpbeutro wtboun the foruwd rector (6 moky a rocante onderd per me perang den 005 phosphate, HI 1425 C) bartetnih ule us 100 thor the action Dard in the Theme + ud ++ adhat Son of formant recortamord cont cartabon bd fant nota that u not to mentioned multirety All the nta ondentor cared to be presence of a sortute or 003 fronte Froon Vamry und Seger reproduced by peroplanch of the morm Parlapan Clement OOOOOO bare dappeared, 1 extrapointed mfinit dve oncentration At actual concertntions the i mity may be much lon et Ferrayand barres only about one hall of the actrity of the entrymu and, trnally th revera racion the reduction of fuma note to roccinate with CNH o electroa donor proceede at bere than one In enleth of the rate of the udation of Bucetate When ne ampare these ctrico tb ander data for the soluble delly drogtoere in un centrally borzogeneous stat (TABLE 3) sobe string differ enco become cuideat As erpected from the fact that the cytochranne chum has been removed, the res taons ith O, and clocrom and those with dyos (auch u Deoretruechum) that reect lar aloor the cyto chrome in, are also lost Sunce in particulate preparation ferryanide bus at laut two reaction utes, the chief one being at the level of cytochrome or and the lewer ooc with the dehydrogenake the bolted dehydroplane reacts relatively Ittle nth latxade the reaction of metbyten lur the routine unter and bount crosyt blue dyas that opet krved! detisdropeas in thes, with the dehydropowe bo negliglie Am muy burtowe therefore that they mat en here love the best the the hydrogenate In the reverse rection, ce tron donors buha i locity with the FAD moiety of the debydrogenate, retard the foll a listy on more The post surprising fact in the table bilization and holatron of the note ·488 Annals Nery Hork Academy of Saincts to that, u compared with FINA, u electros donor even the activity with phenazine methosulfate had been reduced to about one hell efter solation of the earynie. As we nou know this is because phenazine methosulfate ako how to reaction das in the then sport chala, one with the dehydrogeoste us He know it add one slightly farther along the chan the latter or lost i hen the dehydrogenese bolated It may be of interat to Dutnte some of the sechaiques by which hxh conclusoos may be reached In mitochondru the oddation of altimate by peotetrolion and related dyer is anumycin-sensitive bence their action ute abould be abort the level of cytochronne b In fact there some evdence that cytorbromx andare is the ute of the reaction Similarly in such a system a major part of the ruccinate-fermeye de recuona inhibited by antimyco thuit mould dearty be cipected that a major part of the Arctinate-ferricyande reaction would be lost in spanting fanc dehydrogenase from the cytochronne chain Thib, in fact, the air Another techniques to compert various types of prepun uomol the electron transport chain and of fragments thereof all of them apable of ooding succinate at high rates, with rtyand to their relative ractiyua with vunom electron scceptons The venous type of partkulate parcanic arkdare and fuccinic dehydrogenase preparator in TABLE 4 have been claimed to have very different dye specificities, one of them reactlog only with brilhant crery! blue but not with methylene blue, other only sith ach substances u Indo phenol dyes Such contention seemed unlikely face the rencloa ates in the cytochrome cham could hardly have equaled in number the dulerent preparatiom clauned to have sngular odividual specificaties. In fact, auch group of travestigators bad its on fa ante preparation endoned with a specul Danse and for those not working directly in the field, the stuntion must have meemed womewhat chaotic. It appeared quite probable that many of the differences were reflectors of the different any techniques used by mdividual worton In fact, most investigaton unayed their prepuntrom at i pogle arbitrary dyt concentntion blat may bave been so low for one dye that it Tur 4 RELATIVE ACTIVE or VA O PADLA MOUATION POU A IXAL பரH DICயுறு Ram Mutsy (V) Turit th Hetilen hre Che Tarte The 24 100 100 18 54 18 13 28 6 Kdlip Hutite Bull Cooper uccroc audio Al Cooper deorschott inct (tretoSDXC Stotram 1 (cholt tratal) 11 100 100 90 77 15 10 10 100 20 . 03 19 - Alain tia are i.. luas at 3C PH 76 ·t n Singer Electr n Trangort in Cardiac Tissue 489 masured ooly a fraction of 1 rity and to high for other dye that the toyme was almost compki h buted by it. As me base dikurd clue bere, reliable turbon th otidiang mama may be derived only from maumal activities ar i mlated infinite dve concentntion based on parchetk dita. The nyme prufalatkom u TIDUI + Inxlule some that pass the complete electron transport ! won that contain only cytochrome and c and mode that although illy Late ure bargely devoid of cytochrones In oar laboratory Gruditif rib taking the activity with phenadne metboplatc u 100 txert th dyes hore reaction site is close to the dehydrogenuke (meth | Inolunt cresyl blue FIVH) b the te in all the preparation it pton sich us cytochrome c or cotetrazohum, the activity nouk! Large figure to 0 and with clectron anions such us fern.yarude nu! H the activity drops oil in prepen toon in hich the cytochromi h From this type of data and from our knom ledge of the tox hrum 001 t of the different preparations we may deduce which electron atent act lose to the d hydrogeous and which have reaction to far ber removed from it A very different technique for the emploration of reaction sta in the clectron transport chain us the use of differential mactivation Lor concentrations a cyaarde of course nota taneously inhibit cytochronne oddaxe As Lnong from the word of Trou bigber oncentrations of cyanide moro skoraty Lnactivate the ondation of sux Cout with crtain accepton in mccfnk ddare preparations As a rerult of stemuta study of thu phenomenon, Grandatum has nown that when th cyande 10 nations omplete of the reactivity of the dehydrogerie uitb roethylene blue and brillant cresyl blue sa kort but only 50 per cent of the uvity with phenazine methosulfate and about O per cent of the activity with fertugande (corresponding to the antimycin- In ober words, that part of the activity Ierative portion) are bolshed of an ckctron cartier that reflects direct reaction with the debydrogenase X fully retained As espected the cyansie-inactivated crymde rull recto manually with FUTVH Tow activation by cyan de converts a highly ope that beha as much be the solated pnorganlıed respintory system Bury dehydrogenase One reason for crophwang the plan of the ork on the precinke ouds synet at point out some of its ampoules and the consequent danger in deriyut coochosons bout the actity of the system in poroul or putbological mumple without due orickmation of the dequacy of the way method Dulorentaal inactivations and mhub thors in one may but not in iother are Such frequently encountered in nort to the dectron transport suena effects often occur wpontaneously and may lead t artllat tous results ID 10lmal tbt coatinuous supply of energy derived from Tobac processes, on beurt the properuos of Riktunic debs drogenese meet the requtrement excellently in that ibe formerd a couple There another point of partxuler interest t biologuts whose phypokopal functioning depends on Fielding reactio (cenat oodaton) predominates by far over the rest ·490 Annals New York Aademy of Sciences To 5 RUATIY VA ar DUTEUAL Surtdoc DualDOCUWW . l Arron Cunun kot ) Lectram. Lechat Pro OY1 100 IDEOLODC pretborullat Jabylene Woo Lacethyl pologen NINH, Ferrymarche 1900 125 152 100 62 13 13 100 Lot 900 141 Sw + F Ratio FE 3/1 Uapoblabel data of F J 8 Lan, IGPJ Warring, and T P Spor ar reductive, reaction. The name a true of other cells with amilar toetabobo requirements, as, for example, acrobic yeurs In contrust, in obligate annerobes tho cyme catalyze the reductive reaction about thirty time furter than the oxdative one in the pure fate in whole cells its ability to oudite Puccinate a further depressed by the presence of an mhibitors that Kectively Inhibits accinate ondation without affecting fumente reduction to that fumante reduction predominates one hundred times over wcinate ondation In facultativo Imacrobes, tuch as Propionibadons and P dans bich reqmre the oudative process under acrobu cooditions and must also be abl to duce fumante under anaerobec conditions, the properties of the entyme ac ugun crcellently suited to the phywologkal needs, unce the forward and reverse reactions fuoction approximately equally rell (TABLE 5) Turning to the rest of the dectron transport sykem ne find that the information on its other components a considerably more limited Only tyto chrome c has been solated in pure state from animal tanja Cytochrome type proteins have been obtained in highly punified soluble form from microorganlamy and from live MATOOD, but the wolawon of the denial cytochromeb of the respintory ctuin of animal tones bas not yet been a complwbed Lie cytochrome 8 from heart, cytochrome ( ad « und my (the latter two together kdord u cytochrome andur) have beoo oblamed only u partially punfed partxle fractions or as bile walt preparations Ia view of the back of highly purfio preparations and of specific and for the mensurement of the atalytic activities, it is not furpring that informed ton on their chemotry and properties s very limited. Tith the advent of methods for the purification of pubmitochoodrul per ticles and of their lipoprotein constituents, recent yeu bure witpened onalderable race in the wolation of vanou negments of the cytochronne chain in several laboratorios SCHE 7 talc from Green and his associates, bu dagnomat representation of shat happens when a preparation of the cature coa that unttm Jurnefeltd reported the station of tochron 4 Wlable parroton ·Singer Electron Transport in Cardiac Tissue HO 7 Fan тъ LUCTE I Lavror Cum 491 doc D DPNH D- (Cyt) (Cht) CTP DPTI D Suco (DPWH) debrydrogenas complex SDC ( tochrome oudere 1 The latter contams only It can be degraded Thes From Green produced by person of the leaders Preu lac electron transport hain, hure denoted a ETP scmposed to the action of deoryebobate or bolate in the preek of obutanol or try on The ornare chain s broken mtotno trpent, green particle with cytochronne oxidase Activity and a red one here referred ! SDC cytorturom.es and c (bere referred to by i arlier name cytochrome) batter fragment react with yiochrome but not with further by dearcation with acetone follored bi alkaline extraction to yield an Active preparation of winx dehydrogenate and an macture bemoprotein rendue contuning cytochromes i and In addition to bemoprotein, purified preparations of the electron transport Type contan consderable amounts of non-bemin metallones (Fe and Cu) and as much as 40-60 per ent bpuds Among the lipade a vast number of individual compounds have been recognised ranging from Deutral (atoto fat oluble vitammu A number of yours 40 u oprosted by Ball and Cooper that the fune tion of the bipada u 10 censent the madividual enzymes m the hain to catch In more recent time soon other in a highly recom. onfiguration Thus Nason and Lochmann wecfic additional functions have been proposed have found that in particulat preparatxoo from animal times the show or und between DPWH and clectrons between accurate and cytochrome ytochrome interrupted when the enzyme preparations repeatedly The preparation extracted with Antablo organic sobrett, och u bo-octane could be reactivated by the readee obtained upon coocentration of the a Tuon and Letmann o aboned tracting solvent and by pare nitamin E certain amount of vitamin E from the prepara that the extraction reOYOS They werested that stamm E acts o colector ponebly directly Latet undergoing onda tron-reduction, betutto cytochromas ibere Norhers, as well as other favestigators found that reacth ton of the more ·192 Annals der Vork fademy of Sciences T 16 ICT or Q 775 O STUDY Nutrent Ashtine fortn by an 25 Tone lor More Q775, 003 tochrome 16 Ctrded Letreted litructed Q 775, 0088 & tochronne 0 + 17 29 From Crunc ad reproduced by petoon of the H B Seder Copy ctncted preparation could be even more ela lively accomplohed by a variety of neutral fats and other lipids, including alcomarganne Because of the unspecifiaty of this reactivation and the fact that only about 10 per cent of the vitamin E content of the prepantom was remon cd by solvent estructura resulting in Umost complete inactivation, Slater and others have questoned the role of vitamin E in the electron transport system * The da neverthelem deserva metlow attention but the pbenomenon us bygbily coraples, es never thing else in this field, and its conclusive demonstrations fnught mth expert mental difficult Thus, more recently Donaldson and Nakon have been able to show that in ertain aged prepuntos reactivation of solveot-ettncted mmpla la brought about quite perlfically by vitamin E and suggested that the explanation for the unspecific reactivations observed may be the various lipids move the vitunin E sull repalning in etncted emplos to the active ate where it then functions Among the lipll compound atn ted from heart prvommes by bo-octane and similar wolvents si quinone of us yet undetermined structure to which Iti dAcuvan, Inned drefer u l-275 The quinone sad to be much more oneninted in panfied preparations of particulate nurain dehydrogenan than in the original names. Prepantone of the clectron transport chain are apable of oodlang and reducing it this, in itself, Dot a good cnterton of physiological activity but more uçoificantly it bas been reported to be wolable in the ordered form when the propantins ut attracted after er poure to ur but in the rowed form when they are entrated folloning et posure to cyınde aod succurate As xen n TABLE Q, bart preparations in which wuccinic oddaxe uctivity has been abolobed by solvent ettnction are fully reactivated by 1 pall umogot ol Q-275 but not by other lats tested, lochudoy vitamin E The fotarpretation of their observations ha agus como phated by the fact that beddes the quinode cytochrome c also reactio at low levels, despite the fact that cytochrome co not extracted by the sot veata used however the lateret of the observaLOTS x updenuble Another bughly boterestiny obrnition whose full significance cannot be arched at this time has been reported recently from the aude laborator)" QITS (otoquox) b qurth data th oinone pronously boltolu Norteos tortory For more real godes of its metabobc rok kt datere ·Singer Election Tran port in Cardiac Tiosuc 103 Tuu7 Roca 110 o pritt OTID UND JO OCT rm LCTD IT T CO DTH DAH mttt PH nen + dir inte Tore 3 O 10 75 0 1 Ot 20 O JS 00 Ois 00 00 Therectombe babolawd fraction in thecol or 002 per contestochrome C added valed 'DP 33 modex Hboot An actmtky pro da DPH arterd/pu prolet Front dner and Crabe reprodonar la pared Bwch Dwal Ach make According to tbb report following thobol (roximeat of Aroma lind board cytochrome that oluble in noapour solvent but not la ater may br otructed by a tane The on Ipad cytochrome lormed from ordinary cyrobnome upon add ison of phosphorqud fra 110cm solated from the ASCOLOM Whereu mahrole trueted artin ondas paregama Lorish u dos shown are fully red 1Mied by other Q.75 or ordinary como chrome c in regard to the unit cola brota reut trop the DP\H-andung bully of the Ame emple rooly algbtly restored by ord sury ylochrome bass fully racuvued by Hiperd ytox turome (f4u7) The pooling apat of the report # tluat above ytochrome bere the factor propumed to Kt, the pathway of electmn innsport generally thought to be a common Obe for Racerte und DPH andatan Before leaving the object mention should be made of the role of lumia m the chain us postulated by Various and Vit Lutron (KTLUES) who Hate that vitamin K would act bet cen DM i and cytochromend tour nould not play direct rokem pocrinat ondo hot ever they poder nomia K to be on the normal pathway for DPNH oration beatbe Lattet coupled to the geomaton of ATP while the ytochrome reductum pathway would be doophophorylating bypu The contex on nhach tho shark rats a direct, and altempu io reprat Colpo Boarin und some of th key Obertvitons have been traandul Slato ha shown that turun Ks at ydy ondead and reduced by na lour of VTK Hima, m CILATEOW TL CEAU ACO to Wat Hனாற Cat Boonma Slama Stor de montre parte Voz LDC - tor C - אין - سم aroprocess heproduced by pet utad De la ·491 Annals New York Academy of Sciences spinntary chaln preparations with attendant ordutive phosphorylation This, in itself only compatible with physiological role but in no way prored it, once a large number of substancas unrelated to physiologlal procesas belaire dmilarly an active participation of vitamin K in the chain rencana a part bellty but so good edence bus us yet been found to rapport it. As may be surmoked from the foregoing, investigations of the constitution od mechanam of action of the electron transport system are in a vigorous Aate with a multiplacity of moerplained observations amoting cyploration I feel that clar-cut im to many of these problems sal hare to ENALE the prolation ol the individual eftiymatk components in a high state of purity the development of specific ways for each and ponably the reconstruction of the wbole from its compacta. DPI1 ondation In heurt the ondatron o DPVH, like that of Riccinate, bomociated with Ricosomes and b modnted by a cytochrome reducing dehydrogenaxe. Catam important differecos cst, hon ever between the events m succinate and in DPH oddation While the metal reaction on the axante perb is an ambiguous one between exccinate and its debydrogenase, the imuual events in DPNH onda Hon are more compler About 20 yan 45o Straubi solated from beurt a favoprotein DPNH duphoruse, whuch catalyzed the oddation of DPNH by dyes but not by cytochroma Aboat 10 yain later Jlzhler d alwolated another mitochondral flavoprotem from the same source, DPVH-cytochrome redactus, which catalyzed DPH ondation by cytochrome c while pourendng dapbon activity nu distingumbed from Strubs doaphorixe by virtue of a much bugher Donhemm iron content (4 atoms per mole, os ID RUCCIe dehydrgeoux) Sorao yern Leter Jlahler and Elonet proposed on the boom of some indirect but clever eveniments, that the mon components in the reductas participate in detroo transport to cytochrome and that the diaphoris, being largely devod of these, cannot reduce cyto chromes and la therefore a degradation product of the reductes Subecquent experimenti by Nancy removed the crperimental beds for umumlag that iron bas en ondareductive role in this coryme (although the still romansa powsibility) The lon nevertheless eppaan Decorary for the binding of cyto churonse c by the soluble redocture, but the phytologiaal vigoufiance of the fact 1 uncettun doce, in the electron transport cham Dot cytochrootte but cytochrome c (or pondbly b) is the first herpoprotein to be redaced by DPNIL The contention that daphorase is an artifact of the solution procedare thos a degraded cytochrode reductox, s mal ance (1) the procedure for the solubiustion of both myma fro. Bartosones are drastk uod very smi- Lar (2) dlaphorum bu been abonn to eat to intact drculating reticulocytes, od (3) recently un important physiological function bas been propone for dlaphorise Before decoming this function se most consider hon DPWH generated by pyridine pocheotide enzyme inside and outside bean mitochondria gets to Very monthy Blanche de boastrated coetandy that dapbone bono made ortodoxe reductie behes shox abo that dupboru xt e) potat book od dehydratex Thos entyme ·A link between Singer Electron Transport in Carduc Tissue 495 the dety drogeras, which reoudve it and thus starts of the chain of events chery generation One posbury that DPVT, being in mort ausa dissociable mobile COCTITIO urus of the pyridine nucleotide debydrogenate and somebom reaches the struatus is bound DPVH ordining ryuter of the RICO 1 moment's nk tmn bong to miod come difficulties with tbo mechanism Externalls acted DPVH certain oudised by bear butochondrta with consequunt & sur ton of ATP As has been known for mamy jam, mitochoadna contine Diderable quantities of lightly boud DP that cannot be remon cu mnr peated washing According to Lastera beart tailochoodma onks that part of DPVH oddation, which iluunted by nas of bound DPX o mupted i ordaine pbogbory lethon If this is the cix, bound DP might an Ink been cer the electron transport system and the dehydrogenato e img at carne dostance from it free and bound DP\ pron nded by the obseriebons of Weber and Kaptant that mes eral Aurdenzymes, notably heart dapboru, atalyze otidoreductions between DPWH and its analogues od there workers have in fact, suggested that electron transport between pyndine nockoudes might be a physiological fonction of diaphoraxe Biltochondra and more or low miut preparation of the dectron tranport chain evidly otidie DPVRth), s acceptor ber cytochrome a pronad cooponent of the chan, ben tetoally added, tapes of clectroos only slightly In sich a ryuten und is redexed forumlarly only slowly ouidized (TALE 8) Preparatou nith purch baracters have been termed chored by Green and bis co-maler," opposed to open DPH ondare preparation in abich (a poor and lerncy tochrome e a good, Acceptor from DPU und The terminology for which ferrocytochrome s odly oudered by On meant to upły that be opening phenomenon mterrupts a chowed cooductor yre u are one ut Opening of a cloud preparation readily brought aboat by deorycholate or trypen traatment or by (VRSO and coolate The latter tratment in lact, has been abonn to cause sepantom of the partecke TABLE $t Dom Outs TLCROLUXATXO CALTTC Prim or DMR DHE TPL DE pak uthy Dr C) red gur OnMout En Ear Dre + denot) cholest KO 00 Martott nu HOM + Varech at 50 From Gerakan produced by perros d Bertrek . Bwan Lake ·196 Aanals New York Academy of Sciences 80 DPNH 60H HL 40+ TPNH DPN 20F ENZ CONTROL 0 30 60 90 MINUTES FICHE 1 The oudation of DPVII und TPVA Iach crpetascotal Hubert card Cootamech, tibe Hart, 20 ml 01 PO boller, H 74,01 mol a tochrome 6X 10-11 & mg (upprunmatety 10 model of DPNH or TPVH and HO After 10 DID 01 ml of beurt macke prepuna bon apped from one che art T the rol coolant TPIL, O 5 mg o DPN (01 nl of 11,0) u tpped from scood de un after 40 mm Tbo final olume un auch vend 30 m) The control al contamed no TTNH or PPYH, but all other componente, anchading 03 mg o DP Fron Dalland Cooper into two derivatives, one with the characterstics of cytochrome oodax and the other with more of the properties of DPH-cytochrode reductame in that I atalyzes the oddation of DPH by cytochrome c* There are two out Handing differences between the cytochrome reducing particle obtuned by thu fagmentation and Mahler penfied DP\H-cytochrome reductase. The latter devord of hemoproteins and anumycin-okengtive the former onuns bemis, hich are reduced by DPNH, and the opdavon of DPNTI by this preparation antimycin-native Are we then to usume that there are two karate pathway for DP\H oudaton m mrtonosna, the antimycia thotne electron trumport chain and the DINH cytochrome reductes or even morte, that the latter a notber urtifact al polation? Not at all It budl to naall that the ability to reduce cytochrome C nprdly La bad chemical fature of flavine, retured and centuated in the flavoprotein DPVH-cytochrome reducture. The redactes out well be the inital dehydrogenase in the chun of DPVH ondation, pues Ing electroos on through hemoproter(s) to cytochrooxc When polubulud und polated from the rest of the go to hrome Guin it may xil retain its ablity to reduce the ytochrome composent with abkh it normally reacts (the point has De er been tested since pure preparations of ghe Dot been obtaloed) but because of structural umilanty in the actise center of how proteins, it c n also lol and react with cytochromec TPI adalint The best pobject I should like t touch upon briely ·497 Singer Electron Transport in Cardiac Tissue VINE 9 DODORED TO of TTYNALDUAL TINN Sabant DEVRA O con Sotente - IPVIL USD A due to the po 1 th me Kutana rewalmall pameran Bahasa Baptric cun the mechapun of TP\Hilarin Intimportance to arduc entry foder atroa ban in the fut that one of ih kretank corymes B TP\-specific and TPV balso the probetx group of other dehydrogtes of Importance in bart metaboluson The ondt phophytion accompany ТРАң oddarson in heart mutochrondre and to be moch leus ettenerve thon is the aum with DP\A La fat pertolk painted preparations of the electron nnport ritem from heart annet en ouduze TTVH Able rechanom o TPVH oodation and tompunging phosphorylation in beurt case from the work of Kaplan and ho collagos ho have detron Poted that the enzyme pyndin nucleord transhydrogenes which at myten ondoreduccions between pyndine nucleotide conIYTOAL, prekut m Total un mitochondre FIG I reproduced from Bell and Cooper * pro ndar evidence for the more of the irupsbydrogmere roution in TT"\Horidation by peopentons of the clectron insport rateta of baan The paparutions oudro DPV but not TEVA anda atalyer mount of DPYintata ihr oudh tion of TPH Scarve 9 prevent Kaplans xdens I the sequence of foot in oudoreduktoru af TPV mn animal (o The TP redaced by dehydrogrouxen interstis nth DPV na transhydrogenant regenerating TTK and forming DPH The better b then indured me tbe DPNH taxe path wiy muh the concomiant CIKTION / ATP A part of the TPVH mape this route und erre a redu.tre process, such as latly and wnthess Thue transhydrogenase would sowoc a possibile role here ut potential regulator of TINң аbоlага Codwin Ranks The fare cont description cenfetchy and perhaps a amplified, de mcmiben the state of our Loonledre of electron transport mechanine u bent As may be seen, the problems ire amples and the competenta duliculte condarable Itu al to be aware of the uncertainties, difcutties, and meny pullalle on the fickt, parakulerty one * to embark on a westigation of pathological changes the tem . hihnay wuk retaurely len roude The theatru transport sitema Some of the bacba o crea but ith many branches that keed of it an bypass in that oddation aran wibout any conservation of coergy (auty there are many was bich the main puth may be blocked ite demonstration of the presence of the cotopopent tnyttet i putbol ·498 Annals New York Academy of Sciences sample is no indication of the functioning of the overall system. Furthermore, 1 manurement of the ntc of removal of ouditable mubaratas, and to of total O, uptahs, gives no indication of how much of the ordative energy mu CORKTYed in the form of ATT The powubilitla of metabolic derangements are numerous many of them may lead to wow.oupling of oxidation and phos phorylation From the standpoint of localiang pathological changes, the efficiency of the over-all functionlog of the energy-geoerating system open to and Hipe for systematic study even though the monato details of the mechinery are pot yet clearly defined Asyatemat comparison of the nata a oxidation of mouse key metabolita end of the efficiency of the accompanying phosphory! ations in marcosoma from normal and pathological hartı xems to be highly dourable. 1934 Rearance 1 BG, RJ 195.5 I Cardovicolar Surgery 2 11 CR Lam, Ed Semden Pudephon, the 2 Fucit, CK 1956 Dua o the fort. 12+199 Zod od Sam. PLI acelo.hu, me 3 ETXEIT & F L CW 1956 Ilorin Nitrogen etabolum : 601 WD Elroy & Olu, Eds Johns Hopkome Prem Baltimore, jid 4 KX, DET HARLO 19.10 Proc Boy Soc Landon B18 277 3 G, DE 1956 Intet abortal Sympoam Emry ma Uputs of Blolgraal Stractare und Tunction : 265 OH Gecbler Ed Acidente Prem New Yort, NY 6 SLATII, EC 1919 Pocbom J 4 14 7 REIT AER POTID Arch Brocbom Biophy 48:1 & SOOD, TPLB KLADNY &P Bath 1956 J Biol Chem 999 9 8pian TPLB KLAL & Vin 1957 Adrencan Errymol 18 as 10 Soon T P&V Jamn 1957 Record Chem Prog Kreat Hooker Sa Lab 1.: 169 11 SEALEY, LB 1957 J Bal Chem on 303 12 HANIT VTP STOD 1957 J Biol Chem to 753 13 ENA DOO, RP 1957 Federation Proc 10t 178 14 OIODITTA A&EB KLAUTY 1958 Federation Froc 17, 220 15 KaxrY, H, RI_GEAD VOITT 1937 Fedonbon Proc 16 2017 16 SIRIPIEB KLAPIT 1957 1 Vethoda d Brocberakal Analytos 41 307 D G , Ed Lotericlect Mer Jart, NI 17 Trou C L 1951 Blacbem j 512 18 MAHIY V &TP Srom 1957 J BI Chron pet 203 19 WAMI, BI GP J OH SIG A GrupoItA & TP Saga 1958 J Brod Chem 90-97 20 OUTULL E, D TAOTLA, L SEUTU & T HORD 1957 I Abd Latera Sydpodrumo co Edry me Chem z Tokyo and Kyoto, Japan 21 TAILAZA H 1951 J Brocken Taky 1 n STRTTKAITO, P&J I Yuck 1956 J Bal Chem 1 200 22. JITELJH TATKALA & D E G 1959 Federico Proc IT 219 23 BALL E GO Coorn 1909 J Bal Charm 180. 113 24 NAO, AIR LDXDL 1956 ) Bb Choo 311 26 Dool, DEC SumnEL YILDILA 1958. Biochen at Bopb Acta 17) 13 26 DOA DONO&A Nuo 1957 Proc Natl And Sd 4. 364 76 FPDAO, GNFNuro, J 3 Lon&R A Jato 1955. Blocos 266 SOTO, RAGNOT J S Lon XAVF Lut 1948 Blachan J - 16P 26 m., RLTO ET Raru 1958 Fedoton froc 17s 238 # SUL E C 1950 I Sopoauma V tamin Hiabob, 4th It Corpo Barbro PODDOO Tren Loodan, Fagund L proce Joh 58 ·Singer Electrop Trongort in Cardiac Tissue 499 na FLI HITTIR LLORA IC Haun 1937 Blocben. at Broply. hete It: 220 #HA, CEFL CLI 1958 Brochure, et klopke, heu 87 203 9 HUS, C. ID ITT Loren 1954 Hodun et Btopky leta La: 152, p Cow BOTTLA, PIEL SLATIS 1938 Barchet Barptyn Acts 122. IL Sran, ID 1200 Borbea #777 $? Vunni HR Suru Iria 1951 Dla Cbetu, 13+ S&S 1 Yunni, RRED G Elon 194 J Bal Chce to 105 4 Sury'i 1957 J Biol Chem wid 33 Rarirout S&C 1957 a phriol Chen Hoppe Seyler 300 196 * m, 1 SI & Carla 1957 J Bal Chem ou 909 * HARRY ! 1958 Brocbien ct Buty Acta th 20 1 Nans, T&DIR 1050 Irch Drochen 57 418 # Lama, R L D Liron LDL cron 1957 Blochilm st Biophy X. 155 Gm, D E B Mwan, R Rar II Rann 1954 Broche et le Acta. 150 LS Acun, B&D Om 1950 Brachine at Bophy Acta L 41 Visus, P&P x 4 1957 Bod Chero bo zos KALAVNOU S VEFraas Corti 1956 Proc. laul Acad sa u. 131 U. KALL, E C&O Coora 1957 Pror at had sd 01. 157 Arta.

·OXIDATIVE MOSPHORYLATION IN VUSCLE SIITOCHONDRIA AND ITS POSSIBLE REGULATION للعطر I Philip Suckevits 1 Rakoto Taiju dla Rocach Xbot As my work has been done on live mitochondrle nther than on hart mucle, it becomes incumbent upod me to compare the work on livet mitochopra with that reported on heart mitochondre, aloo alled arco ona Ondatrve phosphorylation per we will not be covered as, at present the subyert be too abstrux and controverul, und the mechanisms are still sought cmphaste the regulation of oddation through the coupled oodative phosphor yhton system, a procesu rell known in live mitochondra and somcwhat les well known in beart muscle mitochondria und congda bow this regulation might be reflected in the nork of the hart Fint I shall docum brefly the known rudiments of phoxphorylation as coupled to oberate ondation and to electron transport The sequence of reactions in the respiratory chain, u given below us known chiefly through the work of Chance and Wiliams, bile the poulons in the chain where ATP u farmed we knout n pomanly through the periments of Lebninger Substrate DPVH flavoprotan 1 - ATT 2 ATP Cyt. - Cyt c- Cyto – Cyta, - 0 ATP In the case of some rubatrates sich us glutamate or runcinate there o no phos phorylation at the aubetrate - DPH Rep. It s definitely known that the oddation of reduced cytochrome c by orygen giver ooc phosphorylation * bile the oudation of reduced DPN by cytochrome given tro phosphoryla thon, ope probably between DPNH and flavoprotein and the other probably between cytochromos bando In all cases where the pyndine nucleotides participate in the clectron transport chain three pamphorylations occur dunng the pange of dectroas tomard orygen 1bb 1 all that rus koos n about coupled phoxpborylation Recept do coveries, however have given us more inoght Into these ra troms found that if Inorganic phoxphate was labeled with and there mus ATT in the medium with the mitochondru, un exchange of the ndoactivity between the organic phoephate and the entenlied high-energy phosphate of ATP touted this occurred in the absence of substrate Furthermore ADP labeled la ita paride mosely with Cº prve un axchange Taucton ith ATP which then becunc labeled in its purme molety Since there reactions are reversible and ene po pet electron transport occurs, ji su postolated that there etats * thun the mutor bondre a high enero compound that oulu When ATT reacts with a poolulated unl non n component The ret trong belor pt un adequate up-to-the-moment explanation of the receot cxpurt menul finding, with PX being the postulated Intermediate. 800 ·Sichevitz Ondative Phosphon lation 501 ATP + P ID + PV (2) It a thought that reacts with smo ober(s) of the electron transport bain abone according to pelti IT Then, during ricrible electron ts ny A+ BH VIL - I+B At the present AH - 4H + (5) Ranction 2 by lud could tb ADI 11 muhange whileractions and 3 could give the P ATP exchange It has been thought for a long time that quinond structures, u ciempfied by the faytos, could not only stuck tron urers but, becaue of thdr Hrtkcture could also parta pat la phoghate esterification me therefore, a geat attempts being med by brou hemsis to deally the component postulated bove I oncian #bether it ompromet ade compound or three different compounds, ode involved at och ute where ATT larmed betbet ut som quinord compound woh u vitamin K or even bether it might be adentkal uuh the devtus directly involved in clectron transport. Brefly to the present fate of the bartochondrial ndetive problem I believe that the whole mechanism of andative showpborylation will be maded oat very WOOD Honesty at our present Hute of knowledge, I think it would be more fruitful t descu a higher order of organization, rudely the regulation of the otdative rate of muochoodne that is brought about by the light cooplong between substrate ouderdoa, dertron transport, od pbomplante autenbcatroo The tight couplung vu final noted in biver mitochood by Kelley and Kihelkeyi aod by Lardy and Bellman Toeke worto noted that truth mitochondre ab bited very little ATPase activity and very little oodat ve tvity even when they were presented sub sobitat Bigt und ATP When Borne obstances that broke down the ATT ere dded bunt ad oodatiye Lctivity wus noted From the core of tbe pobstances dded at ni deduced that the Oromoo koomu*th proxence of or the production of a phophate acceptor ADP ( tbx ompound nere present, one could obtain ATT formation due to pubertnt oudeton lo ther nords, lo the frozb Tutochondri, ADP forma too mat arus in order to achei oodatna ai tubante TABLE 1 prekatome data on the potot. The compound DVP Down to break dona ATP by some yet unkoond mitocboodrul mech ungu In freab mitocbondrs there is no break dorna ATT OTO In the Thos being prokie of 318t+ open a compound auch u DVP & added medus containing ATP od all the requiremkou the tk, mitochondre, in for ouders, will beve rory little repertoa onko the ATP a broken down to ADP t prorride an atoptor for the borgenk p bocephate during subent ·502 Annals New York Academy of Sciences TAL 1 Im OT DNT O RATTI GLOLAKATE ROMO Y Lin Jako Dk7 Coptelen 0 J X 10 15 x 10 J X 10 15 X 10 00 1.30 350 160 mo AD the thesis contumed pbophate bufe tochrome MC ATT und poterat m ppropret apocab Keprodoctd by potteloa of The Joy of Biological Chemistry podation. Not only will added DNP have this afect if berokinese mod glucose are added, glucose-6-phosphate and ADP are formed and respiration IDCCXL. If creatine and cratine kinek are added, pborphocreatine add ADP ere formed and respiration ugain untuk By setting up the appropriate conditions, Lardy and fellman' also showed that the concentration of morgan phosphate could be the limitlog condition this concentnton fell beku e certen level, even though esough ADP were present, respinto leveled od because no ATP could be formed Tho berokines and creatine kloux syntes are citrancous to the mitocbooda the DNP system , in affect, an artificial one Potter and I there fore demonstrated that syntbex events in the mitochondra, uning ATP could increase the mitochondrial respintory nute TABLJ 2 shows archi result To the medium contaming the substrate and ATP he added the other necemory components of the citrulline synthesiung tyttem, the respons ble enxyrue being found inside liner mitochondria Il to apparent that wlule this tyutem did not give u hugh respetary DCIELE as did berokimas substantal meter mes nevathden, am obecrved TABLE 2 to Rows the Interesting observation that the herokinase added to the medium could ompete with the atrulline-fyntbehang syuten mithin the mitocbondria for the ATP bang formed within the mitochondre. He also wanted to find out what happened to the ATP that wus bedog broken down by the herokmase or by DNP during the codore of the stimola TAL 2 STIMULATION BY COLD SYAD 01 OODATIOX X LI WITOOTODALA Varun On puke 0 J 13 40 120 65 Non lleolinux, cocore buyaume, andt, Nha Tomkiner, racoes, orthoc, NHC arbyl platurate 0 100 AD the coursed ptophete bote VECI, I tochroare s ATP founts, ed py the propelut imoonts Reprodend by petebat and Burgic Cat ·Sicheritz Oudative Phosphorylation Twu) FIT OF HOME AND OF DAP Oria Urtur La STOCD AND Ara SOTTO' BALAKI Nr Vorox 503 AT hub RUNT ADR ht 35 Voos 00 L break 004 al baratte 006 ml betolibre 000 m herol beer 26 26 16 11 08 04 04 04 7) 77 Noor 17 0.3 X 10 DVT 38 07 03 X 10 - 11 DYP 40 02 10 X 104 V DYP ot 04 43 X 10 V DYP 61 02 24 TEL All the Bests contased poopbat beder och cytacter ATP od ratanata ar ketoglatert the propeat mots Reproduced by remod of the Jew Blog Chow NAOOO — TEILT thon o oddation by these agents TAB 3 gives the interating and me what mirprung regalo It us be men that u the DNP and betaline vitems are loctered, the ondation of puberto o microured prepunably doc to faster rate of break down of ATP Horever ap to cutain level of Bodhton of heraliranse or of DNP the oxidative rate and the promphorytettye rate of the mitochondru cunkcep pace and thus continpously reuynthetido the ATP from ADP and inorganic pbocphate us far as its being broken down by the added agents I think that thche apendicato dondoostrite within wide Imits, the mitochondru can respond to noured loads or demande spon therm, whether these demanda ere external u in the ause of herokinase or of creatine kimax or intern u in the case of DNP or of the atrulito kyatbeinng systein It could be and that the oddative mechanism of molto doodra is reponave to conmanental stomwons and that this rapoane te regulated by means of the loghat coupling betr een oodation and phosphorylan tion for it tuppens that the product of cudative phosphorylation to them the mubotratc, no to pak, for syntbetx reactions, and the degradation product of these synthetic reactions, ADP woo the subetrate, to a renes for oodatire pbocphorylation There are other methods by buh live mutochondre can be induced to ulter the metabolic activity One of the fint to be torreguted wu the fact al ultered tomaty Era ance the lata work of Scopeder a with liver mitochondre, many other meallgators have attached the problem There is no doubt of altered metabolk stivity of macke mitochondrta u That the intru iure of the mutochondre bxh could be changed cod mergh by henge the look ty of the medium, hara gent dlett on reopis tory and Ilon ever I shall not calorat on this, Lauke Rhoqhorylit twin tv the record of way obecnatione ue Dot be the physiologa al sgnlficance In bury and in orde colors of physical relevance, such us etperimenu suh Honet I MULT thyroids, the date to the iterature are often contradktory ·504 Andals Net lork Aademy of Sciences present sooo bile and exclumve reports on the diennical macaning of rollo chondrial structure Isolated nat liver muitochondr, when placed in mater swel up this snelog bs rather datinctive, for these membrune-bound structuras do not act like balloons. Water reems to enter the space bete een the two mitochondrial membnnes and a bort while, the outer membrane begins to detuch udf from the inner peobrne at a particular pot on the mitochoodrla surface The matrice of the mitochondria become ladenie a proteins leak out, and the overall mitochondrul dimater reaches two or three times Dormal, due undoubtedly to imbibing water Something similar to this, although not u drosta, happens even m 0 25 I microse solution, partkularly if the tempera ture & devated At the name time, the acid toluble, ultraviolet-aboordmg compounds within the mitochondria begin to lal out of them Together with these compounds, whikbare mostly the edenine nucleotides, ADIP ADP ATP DP and TPVH," the Jig H+ Net and it roas ioltally in the moito chondrta come out Also the ATPexe characterstras of the mitochondrie ut altered from DNP-sensitivity to , 18++ sendtivity and the abuity of the mitochondru to couple phosphorylation to ondation o lovt. Some of these phenomena can be prevented and some can be revered by having the mitocboodria do poole work for cwmple, by ordung Rubstrate und making ATP and in some cases even by adding Digt+ and/or Atin or by adding DPV to the medium Even though the functional melding of these apparent reverubilities i not clear Ish to emphaare the froportant part that structure plays a rotocbopdriel metabolism Relevant to the entire documon of the manufacture of ATP the following facts are known that liver mitocbondra contain ATP that this ATP » bound for it an be released only after the mitochondria are damaged that the phosphates of the ATP are in equilibrium with the phombates of inorgaox pborplate and of ATT-phosphate during oxidative phosphorylation and that during coupled pbosphorylatson the ATP manufactured by the mutacbondria is not stored within the mitochondr, for that mechas to be prekat only a certain nomber of specific bloding mto In other words, the liver mitochondria act us secretory bodies for the ATP they manufacture and since the star of the dectron-thingport entymes and porbly those of the phosphorylative enzymes are located in, or are a part of the mitochoodrual membrane, any aterations of these membrana an have gnificant dels on the manulucture and secretlon of mitochondrial ATT At the present time, hon ever one cannot ay any more than thks, once the equeriments relevant to functional regulation of unsture have pot yet been performed In the remainder of the presentation I shall compare liver and beurt mucke mitochondre and shall try to show that force of the same fators that regulate the metabolism of livet mitochondra can bo be operative in beurt muscle Ditochondra An electron micrograph wad to produx encorn 1 100 2 the batter bon ng nat heart muscle fibers. There remy i be strict locall mation of the stochondna ather at the level the A-lanche ora th I bund Ficutr 2 hors 4 type heart muske mito hoodrton Haut muncle mito coadra are about the mame de and shape us liver outochondria they baie a

·Sickeitz Oluve Phosphon lation
5
Frotu 1
Rotwander (onde)
x 11,400
double membrane surroundmg tber, and abo the Cambar choiuas thet
zgination f the ner membrane
They also contam mult minmtochod
drwigragukes similart those food on liver mulacboodne the function of tbee
ules randon
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·506
Annals New York Academy of Sciences
-
From 2
Rat mogut (votnck)
x 55,200
tbe muclo mitocbondrk hart Duy more catre than do livet mitochondng,
und Pabeden has poloted out that this might be the bordbokoglal counter
part of the increased terminal respiratory activity of muscle over llver dito
choodie

·507 Sicherits Onidative Phosphorylation Certain other expects of the url on beurt muscle mitocbondre are relevant Plaut uod Plauta found ibat hart mitochondria could onldite a variety of Absenta of the Krebs seke longer cuinte mis not oridized anden DPA va added, and even them a Hoz nate of oxidation orunted. Despite the finding, these workers had good vieme that the Krebs cyck me operating Chand and Slater found that is differentil crotrifugation they could botate partida, which they called as These particles lze livet mito chondria, ontained a coupdktable wount of the cuccink dehydrogenak od cytochrome oddas actives of the shoke muscle bontogenate. Although ub Incloa contained alto high percentage of the eletoglatante oxidase Syreno, It mu found that all of the moschinese or adenylate Lines activity bich is found m threr nitochondru su found, pot bo muscle carcomones, but In the carcoplasma It could be argu i that myok mex, which bully at tracted from livet mitochondre by us lout from the auch MIDWORCA durmg the bolation procedures Slaber aod Pauls soned that beurt mitochoodne could give a good coupled pbosphorylation as does live mitochondra. These worden derson Haled that the one-step oddation of a-ketogloient to sucdoule by puinenple heart mitochondru give a P O ratio of 34 na the aberce of theoriche, wblh bu good us has been obtained with Iner mitochondrm Furthermore the orklation of glutamate, fumarate, malole and a bydroxybutyate save P O ration of at least 25 bereu rumunal ondation are 18. Holton aos obtained simular values trow rat beart moltochondo, concept that they buahned exceptionally tow P O nato (ten than 1) for maccmate onderhoa. Jfont workers in the field scom to ugree that beart blochondria do not breve un impatred pboxborylativo alfa cocy at least is compared to Hver molto chondria Another mans of malog tha compartion to tat different oddative phonphorylation mhibutors at low concentrations on the solated mutobondria a diferent organ (TV ) In renera! bart munclo mito chondria are lem cusceptible to these mahubeton than are the mitochondria As the concentration of the from threr kudrey bruin iod deletal muscle Inhibitort wu increud, both heart and Aleetal mache mitochondria were TULI EJTECTIO Vao. but I/O Rara or MITOORONDRU OF Vwoo TOD LICIJE Het ke 16 32 39 19 19 1 0 AN 3 Duutroptool Jethylene blot Anunt cel dhe Merpone TLUN Just Nu SOLO 08 10 0 TIDO ketigante, AVT Nr grow 40 the Art coupl ptopbato bodo fond, und bekrone bo Repredexed by parent autem ·503 Annals New York Academy of Sciences Tams LT AD NOCHOTIDO AND HD OTH ROSNATO or Flut SALOON Adorotande Optikuu 13 26 Noos Mope ARP ADIP ADP ADP ATT 1+1+1+1 All the cases contained pboxpbute befter found Mitt rekene, und saturate uprgpolet imounts Reproduced by period of the Boardwxced Jowned more rouletant than the mitochondria from other organs From thu peut botated muscle mitochondre at hardier than kidney liver and briun mitochondru Con rapinatory control, us chibited by the acceptor elected as he bed abowo by liver mitochoodra, also be demonstrated with beurt muncle mitochondna? From the dela in LADLE 3 it would appear that reppiratory control ensus in hart marcosoma, for addition of the adenine nucleouda, with and without the addition of herokimx, lactate the rospiratory rate many fold Chance and Conally and Pucker both using ADP found the same thog 1 two to threefold Doctets in respiration, u had Slater and Hollon * At the meme time incidentally Packers found P O ratio of froon 3 t t lor the orxda tron ofaletoglutarate that agreed with the army findings of Blakey and Maut - Slator and Holton also found that during the oxidative tyntheus of ATP ADP nu a much better phosphate uccuptor than AMP tho finding ugreed with the earlier work of Lindberg and Enter However I have alrady mentioned, thus result could come about from the pomble atraction of myohim from the muscle mitochondria for in livet mitochondria, both nucleondes ut equally as well as phosphate accepi OTA TABLE 6 pre some bguros bernog more directly on the problem of respiratory ontrol It an be seen that the classic addition (inc jo the cur af liver moito TAN 6 RoNLATOK COOL LOLX WITH RA II IT SHOES #0 puan None Horobrers, flores SO 25 23 0 0 DNT IDP *, apd cutout 411 the file contund plophat Leder finde, Ngh IT loopt mounts Regroduced by permarton a The Brochenkel Joanne ·rat or For example, Siekenitz Oxidative Phosphorylation 509 Hoodeti) ol ADP DVP and hewlinase-chucose all are in this cake lour to adotal increase in reputation Another agent or the same problem im olyo the breaidons of ATT by freth Bilanhoodra in the presence oled sitt Liver mitochondru can mot do this banke u mbaratc and inorganic phosphate art added no resptra thon actun buone of the lack of ADP to at a phoghet eruptor Hart milactondra, u zaplented tsy Slater and ho carorleto have higher bol metabolx rai and the reason for this could lic in the obecreations that added 318++ and added D\P both gove equal lacteuses in ATTx activ Sizes with theirt arromanes both are added together further doubling and ehe sale of hydrolysa talos place* Of course another remoon for the higher oudatne nate could be due to the meater abount of the cytochromos iban in the livet mul actondra " The stability of the ATPax acthritses of heart mito bondru parallel those of livet mitochondria, for the D\P afect su not lov ter Boruge for bours at OC and the dirt+acuintable ATP *us not decreuad aiter sorge for deye utbe cok However both livet and heart mitochondra uul bom tbeas DAP activalabile ATP if they are kept at elevated Imperator for short periods of time" Euctly the aume thing happena with both preparation the o-ketoglaterale oxiduc syutem tered tha nyacetn nequly arstable in borbe prepartxoa I heart orosomes allow the TLSYRIK Activity of tax in Akrome 125 C for 10 1 15 mm" Da all these buses the quantitative amilarity beto a liver and heart muscle mitocondre an be obutved readily The follomong otisora may bare some phycological opificance Vitake a su conducted esperuneau much they woduced a malenum deficiency in nu od then waared the pbosphorylation efficretsy of the mitochondria solated from beant, Idrey and net They found that only on the cure out beert mitochondranathe P O ni bor ered, thu unt about are helt that of condescent animals They a50 found that makedows of thyruxisse injected mto normal nus produced a loncok of the Prawo in heart mio chondru, brat not in me nstochondre Both the effects are reretted by the inter top of ma premom Since 11 okpown that conned thyroune injectmn decrease the Krum level of magnesium, these sruults induite (by no moins conduively as the cathar themselves adno ledge) the powable mai han en by which thyrurbo CHR WICTEAK: th metaboter rare That Aby utk ouprong oudsteon from phoghorylation through ao Vight locuted effect regulatory den ke or beide on descono lont ind Axay roxation could occur In these dart of integrated studies, u dot eeproting that andies on the effect of some eat, bach u thyround on mete baacbox function should be paralleled by a periments to stow effects on other permeten la blony It has been found in the are, the morphologral characterista d the cell <but thyrounc aur muried being hou tenkred by opixul dentury Deusredents throat pe o mat It und wdrey mitochondria On the other hand, beurt much butachondria are affetted only slightly It has been found further that vanouo orber sent, touch us digitories, are exter of ·512 Annals New York Academy of Sciences In the latter cax, phoy boctatine the phosphate donor to ADP must be generated by the act of ATP on Caune thu do Det gain of ATP ATUD Of course depending on its capacity tha system acts ut reservoir of ATP and its release of ATP con be ud probably , pbyulologically controlled In the long run, hon ever the tyran can get its supply of ATT only from the mutochondria Therefore it is adent that the mitochondria are the main kources of the ATP necemary for muscle contraction, and that the nate of production of ATP through oxidative phosphorylation can be regulated by the rate of apply of ADP the ADP comung from the ectomyoan system of the mucke ihn! Vluscle working at a low rate fupplxs 1 mall amount of ADP to the mitochondru which will generate only the anall amount of ATT necemury to maintain the low load. If the work of the muscle w increused, more ADP hence more ATP-ul be pupplied enough to mantan tbe in- Cred work load of the mucke This limited pucture of muscle functioning omits many obvious factors but leuds to sevenl conclnions Any damage to the mitochondria wul womedutely affect the functioning of the muscle Any damage to the contraculle fibers that produces low output ADP 1 damage that might be corrected by increasing the input of ATT -Il not be corrected because the amount of ATP controlled, m the long run, by the aoount of ADP produced by tbc fibrillar syztem It is pomble that both the fibnls and the mitochondria become degenerated if they are continuously subjected to a low work load. Iluscle atrophy ulus trates thu, us does the finding, in crperiments with liver mitochooda that under dentical coodilom thake mitocbopdrie that have been allowed to function will last longer than those kept from functioning Thardort damage to one component, the mitocbopdra, will also damage the other component the fibris, ad vice verm. In tune thu damage may becordo UTCveuble, that even if we could somehow supply the damaged fibrils with updamaged mitochondna to act as ATP donors or of we could apply the damaged mito chondria with per fibrils to act us ADP dodon, the fibres w not contact nor will the mitochondra generate ATP Despite thu pemimatk prcture of the follog heart I hope that ths deliber udy exclusve vien pount u have shed some light on the brochetaxaal mechun moms involved and that events are not as revocable u I have nyhed them dchronlagnent I express my appreciation to G E Pabade for his loun of the electron mcrompb med to obtain yours 1 od 2 ROKO 1 B&ORD 1956 Raptory den and cxhetore pboboryka Anaco in Enyo 11 5 1 Lorror AL 1933 Oldatrye pbocpbory than Harry Lectura Ser &: 176 3 DOTA, PD ABFALCONE & RH Hros kartu Ometin doodlit pbophoryho N ture 1741 401 trag 1954 ·21 Lao Potin Siclcvitz Oxidative Phosphorylation 513 HAL, CLEIL Loroa 1958 ATP id echange reaction or otide un pbozbory lato Fadenton Toc 11 99 3 Sun, E.C 1963 Herba obowpborylation to the reptratory cham 1975 & Coom, CUAL Lm 103 1957 The way trtphoptate pbophat cherre rection J Bal Charm Dk 561 1 Kurt HRK kiau 1951 Bly and adenosioc tropbop balak nordatn popor atoe J Biol Chero in 48 8 LAND TAXI TU 1932 Oudtn plegborylations to a morpinc pleaptat ud aceptor cetrol or petabaix rates J Biol Cacmn 1 215 SOITE, PIR POTTA 1983 I tuontochondrial regulation of latn nt J Bal Chaco 2011 10 Hoorn, GIC SCR IGF TALADE 1913 Llation of betat motton Cootra from nat oppe lochanual proper a tocinotru in nhau tro copac pertolat lemal j Bor Choo 17h 619 11 Porta, R&ROR VOEL 1951 Rantation of the nit of obtain to nat er putochowa Po poros letabota 1: 377 W D F bron & B Glam, Ed Jobm Hapim Prom Haltimore Mid 12 SLATTLE CIXI ULUD 1943 Efect of tomaty of the occurrun there patory cod phosphorylatos atnity of beart much wrote 57 Bandera J 33 13 CLILAND KEC SLATE 1953 Thamro of hart punda Creert J Wwcroscop 50 9:37 14 ChaiLL, BASI Por 1934 Biochemical soal ompotx properties of sheltal Drunck otocbostru atare 177 1094 15 Ianu JR KITI ALAR 1935 Rektronskap bet coa tha Xructure, once rearth dy and ITP kunty o mitochodna from Digion whebetal Douche Cel Rourcb #: 411 Fq 16 11 W WLTP Sirryt 1956 Separation and dentation of fructe from wolated mitochoodne. Brophys Brochen (ytor 1.69 17 Son P&R T'UTIN 1955 Iotra ortocboodsal components and wideln I Bad Com 221 18 Pic 1 Fock & RED VID 1956 los compt of atar und motorboarina Brochet j H 754 19 II YU, FF 1931 Ouders poophorylation du crtron timeport Topborn Wetabobo 1 297 # Elron & B Glem Ft Jobs Hospil mm Pred Bakupore, Hd 20 Lromna, O IL Fntn 1964 Lector of ordain phosphoryla ton Latare 173: 103 21 # TE XL Ford 1955 hemen samochonden prescabalhome rub powered to delete the boy ble in Chemo 116 357 2 Stuen IT, P B Portu 1955 Retourth labelung d intruntadhowrn! Doclerde coming ouduin pboubory keto band Chew w 297 23 sur PILTON 1956 Easy Os Mocated perto trochu Deabrus frutico J Baper Jocko C tol 1: 053 24 PALAD, GE Elettrow DTOP mtochondru and other toplom lorturo # Ford Hoop 1 tera poem Fanpo Unets a Bolopal Struc to and Faction 1A. 'lador Pro le lork, N) 25 T'LADT GHFXK PLA 1952 Omdatn eta boto of beert contorbrochu J Biol Che 141 Rautor) polo al boot mach Byo 20 CDIL C suma n LALEY & FIG# I Purt I veido of oodature phosphorshton by bean J Đd an h TIR'C Hour DEVIC &um 1937 10 Brochrom of properta o'tochondre rotated from krer od beurt The deus of or abaton hii 20 DAGLVIS STTU 1956 pbon batice on the borpboko udan BK Kimiks oochoort 3) VATTU FC II Harun bytant to 1951 Kuuoo the outlet Bobro JA SU medios uod cause cook about 17 Wethod for salon the line ni 11 CIUI, BICN CO obrane 100 1950 J. 205 ·514 Annals New York Academy of Scences centration of demontese debat to wack we foloses contratto Nature 1m 1215 12 Pcm, L 1957 Coppied phompborylation ont heart Dode mrcopo Ardu. Brocbom Blopban 10.00 33 LOW, OL EDR 1912 On the Becham d pboxphorylative map toda emitobooth Erpt Cal Roarch 209 34 SLAD, ECEK CHLAD 1953 Elfect of all on the rupintory and phopbor, lative attito dbortmund wrtowomo Biochem 561 566 15 VITALIJJ I NAKANTRA & D I HOSTED 1957 Elect onunahm der clercy on oddattre pbaphorylation J Bal Chem . 573 3 TARLEY DFC Coona LAL LODY 1955 Action a thyromme on pilo curandra ipd date phosphorylation Bochilo et Bobby Acta 151 991 37 TAPISY DŁC Cooro 1958 Actre a thyronne ording of mitochondre bolated from Hamt tipa Vator 111 1119 » Coora, C&DI TALLIY 1957 Ssd d mitochoochita molated from hamt toa Bochum a Bophy Acte 126 19 Tarn DF C Coord 1956 Efect of thyroape und related compounds a ockore phosphorylation J Biol Chem. 411 #0 Promw, DCEA A LAXDY 1055 Further social on the potuen qurt matad mitochondre Bochim ct Blophy Act 16 467 11 - KORT RW EN Macho G V GLAXAX 1954 Potentan los abu atsoa d repution by bart mondo mitochondre J Biol Chem 2091 151 2 TDC IY OD 2 Con, J I Wray kHR BLoos 1938 Electrolyt dung hat moclo duroy umaboa no J Phymal 1 595 13 TAXO, HSV RotoTUAL 1943 Electrots to change o toorpuqort shock Pabhc Halth Rope US 601 401 H HOTA, AL 19.00 det uod det on electroly to cart rat mede darang molto Am J Phymal 131 440 S Mun H C < D C bom 1941 Relation of cum ud mod clectrolyte particularly potu mora to oluntary artrose Am J Phyul 102: 301 46 STAWIE, JOŁU HITT 1956 Structure and function of toc updo ACarul ble beoorrancsbock Proc Soc Exptl Biol lid 1 641 ·OXIDATIVE PHOSPHORYLATION STUDIES IN NORDLAL AND EXPERLIENTALLY PRODUCED CONGESTIVE HEART FAILURE IN CUINEA PICS A COMPARISOY GV E. Plant und Depard JL Gertler Na Ya Ururity College & UK, Yar Yuk, As ooc trios to formulate blochemlal ypproach to the heart in fullurt ar to the cifect of certain drup och u digitoamn, one la intrigued by an obucena than mudo muny year ago with the heart-lung preparation From 1 froma Erang shows the effect of lacreding ventricular presure on Tygen COD Rumption and the efficiency of outpat by the heart. At fire (curve 1) the dlastolic volume and the orygen consumption (curve 2) whow a parallel toc With increasing intraventncuar pre bowever the diastolic volume begins to decline. The latteresting fact that while the dautalic volume for declining the oxygen consumption continue to mold almost lineur lanhlon Therdore it uppen that beyond a certun ventikuler pressure tbe efficiency d paberite ouliation decreased At higher prepra there rooms to be . decrease in the efficiency of trader onthablo energy t the muscle If a heurtHung preparation ho tha ante las trented with digitale, thero a rimoto drastolle volume (curve 3) while arypen comumption (curve 4) reruing about the no From thu curve it would appear thuat, m blochemlal temu, d'yltala ,, rtcouphog" system that uncoapted With these considerations in mind we looked for cipemental techniques that would make t pomable to upproach the problem of failure and the cicct of certain drop, ruch as, digital, on a buochemical bers It seemed to us that there would be three general areas were one could look for these effects in the bart first the energy generum miie (osidative phospborylation) Kecond, an cergy transferring tystem, Aich is myoksnes and nucleond dipboopbalinaw and, third, the coergy utilisng system, for crumple, uctomyo In the initul esperiments the yield of opdativ phosphorylation of aardac mitochondra from digitalised normal guina pip vu compared to that of normal untreated murah (TAMI 1) In there espericots we wed three batata, one of which maketoglutante with a malonate block ths pelde onostep ondetion of a keto tutartet We compared mitochoodra from the pormal and the digitalled inimale and found practually so diference Noto abo that th orto ptako s almost the same in the treated ud the untreated animals The me atuation prevul with gutamk ach and with cute y abstrates Tbox t would ypaar that digitabatoa of the normal guinea pe does not land to change in the efficiency of oddative pompborylation and bar do efect on th neto dubente andatoa It comed powable to us that the effect of deltats might shor pm mitochondra from into a congestive beurt falut. However in order to perloin ach aperiment, It *us necesary first dort a melbod for pro AL Plate 15 ·516 Annals ex lorh Academy of Sciences 600F QUAVE 4- 16 CURVE 3 → 24 CURVE 400 az CORSUHPTION 1 PER HOUR PER CENT EFFCENCY ARVE 2 12 JO za 50 75 60 19 200 220 Ko - M WORK PER HOUR Fount 1 Gryph bom my relations (dotted ne) on laurd ootpat produced by enlarcot a rok rolume thout changente o mau urterul preure cocy begins to ful us the emot al dustobe dumbbty are approched Captingo kow represents ayro caption Drulus added at bar a ach carte Reproduced by permohona The Jam Paulo (London) 1918-19 olane 52 TAL 1 OXIDATION OF STUTTS AND PROLIHOR LATICO) CADC MOTOCUODUA TO Non AND DOTTALOD Goru Pey Po Dunl a Ketagtatanto + 001 ut bo 3 Os 3 25 sas 637 625 Glutate Acutet 2 99 2 2 2 99 2 39 Taty nmol poto tortate plea 1 pake rodbom monte ducing favore in anupale Gertler developed excha a purnaal procedure The manfattattoo of the congestive heart future produced in the gulpa pig art Yery onlar to thote observed in human fullure. TABLE 2 shom a comparison of the yields of oddatire pbosphorylation of artire oltochondra from Dormal granen pigs and from animals with the experimentals produced failure. ll en glutamate s used as a substrate, ·Maut & Gertler Oxidative Phoqxhorvlation Studies $17 Taru 2 OTDT or SOM TILLATUS AN tropponunr 1 CAIN lmore im VORNAL ANTALS AND GUIT! Pour Conorm FANTE CE PO C O 3 39 1 010 371 Ecoglutant + 001 polo Dett Gluont -Krogtatant 3 12 2 70 3 21 29 645 3.55 613 35 tbac u poderence in yidd ol Oudative pbogphorylation or in Ribuinte auduon rate between the normal and the operated animal. The name thing = true with betoglutarite us mbante (in the abuence of maloonte) In addition, tbese mitochondria show very little adenodne triphopbata mctivity ben fresh Homever be adepostne inphopbatesc actinty an be developed up to a mummum level by the addition of praded heveb of 2 dinitrophenol There no difference in the degree of development of adenosme tripbophatase In reqrone to inded doses of 2 4-dinitrophenol when mitochondm from normal guina ptg hearts are compared to those from animals to experimental ſulare It would seen from there cuperloents that there no diferent in the properties of mitochondra from normaleod operated inmuals Flowever ed tanthe due tous effect of malonate on aketodutaratı ordatlon with mitochondre from opetite annab Lo fact, from the verge thorn in TAPL2, Cannot tdl that there are many empatments in which there Do Otygen uptake whatsoever with a-ketoglutarate to the presenc of 0005 I malonate We are interested in the effect of malonate prumanly because it the first decreact me bayt ever found between cardiac mitochondria from normal and operated animals However we have to plan too far the phenomenon at the time, and do not know bow it be related to th experienta future In general car mulus se on igreement with those reported by Obon core here in tho monograph The proces d opdative phospborylation (CDCTLY goon doo not seem to be afected in congostre future. What we d pot know however a worthern cases of falure mammals there myght not be deeru o the CDCIU innster systems that follor the generation of ATP I lor onda tive proceso in the energy atdinng saan (coottile eteneata) Expertocats are in progress to let there Hers in tbc animals in fauture Aade from the pomble defects on the enzyoutk lee with the ab Animab in fuluis, 000 soodess there might not be changes in certain pbyt al system, moche permeability References 1916-1919 Hood hart sho 2 PL VI GLUPCRE 17 home of ond u tolering por buto but we mtoboru ima katert punem kar I Gang, YU I perpendica L o {] ·SYNCHROVY OF STRUCTURAL AND FUNCTIOYAL CHANGES IN CARDIAC DIUSCLE BIITOCHONDRIA Loter Packer Emilie Rate Jabun Frodowa Umony Poustenke Polad Ale, Mr. The metabolic factors that influence the contractility dardlac maxle may be influenced by the ocenotic properties of the mitochondru present m the trat Hart harcosomes are the major ate of synthesis for high-energy plor phate boods na beart muscle and there cellular structures have been alloded to us the bonic pump in celha because of the action of aergy och compoundha in the control of onmotic properties Since the work of Ruflaub 1 i thot have been numerous reports that adenoolne telpbospbato (ATP) plus Big long an reverse or prevent the swelling of bolated mitochondria. Swelling and shrunking of mitochondria are also known to be accompanied by change in intrimitochondrial water and sons, by changes in highteattering properties and by functioml changes such as the abllity to carry out caldative pboppboryl ation How does ATP keep mitochondria In the shrunken of contracted state? Since the intromitochondrial regulation of respiratory ntc has been shown to be largely under the control of adenosine diphosphate by the work of Lardy and Welman, Suckets and Potter' Chance and Willam,'' well as us many othen, it seemad pomiblo that adenodne diphosphato (ADP) rather than ATP rus componeble for the control of mitochondrial fructure. Tek hypothem bas been enned by Amultaneously monitoring the structural and functional changes in isolated mitochoodrla with synchrooously recording spectrophotometrc and polarognphuc methode Thus for the reallts of there aperurnents have been found to be pmilar for hourt muscle accomme obtained from seven mammalian and two emphibian species A typral expert manto town in TOURL 1 The methods used urt w follow. The functional panametery mesured are resporation and oddative phosphorylation as shown in the ince obtained with the vibrating platinum microdectrode. The out dation-reduction state of cytochrome O shown in one of the spectrophotometrc truca wus mard by following the specific change due to the orde tion or reduction of respiratory enryme at 430 m. There chungo at 130 mp arc corrected for ponspecfic changes in light aboorption at a seat Vive length, in this aux 410 my wing the dual-bam spectropbotomcto de worbed by Chance' In the lost panameter structural changes were followed by the commonly used practice of maaring turbidity of percentage of light trandon ut i domspecific wavelength 410 m In thu crperiment. Tbex Aructural changes can be observed at any ward length with a dopk-ben technique or lry mamning katterer light at 90° whk h has been done in control tretineti A dareare in optkal density here COSTCODd to an opklation of cytochroce b to tbe 430-410 mm ince and to a seling of the stosoma In the 410 me tract. The slope of the doronard deflection of the platinum cukroeke 518 ·519 Packer Structural and Functional Changes 250uM ADP Resplration Vibrating Platinum Q13 Microelectrode 30 ÝM OZ Que Rot Heart Sarcosomos Plus 4 mM « Ketoglutaralo Cytochromo b (430-410 mylo 0013 to Opticol Density Incroa86 14 Structura 60 BCC 410 mu FOUR 1 trode truce indicate the rate of oryten alliation The nto of reparation ha been akulated in prolos consumed/sec find are written beton the tac The preparation bed hero mas composed of nat beurt ectomomen wolated in 0 32 A1 BUXTON modu coataming 0 001 Al errese u pH 74 The ar-tarated reaction mixture of a 30-ml valame in 1-2 cm. cuvette t contafoed 005 M Alcrose, 001 AT KCL, 002 U phosphate ut PU 13 zod 3 mg murco omal proten At the start of the permeat 4 mla-ketoglutarzte wundled us the oddsoble Rubstrate and 250 I ADP wus added us the phonpuhuto acceptor The extreme Left hand portion of the traca wons the now to be on the active state of responaton alled State 3 which is characterit by the presence of phosphate acceptor and Albatrate the rate la eccomenda by npud repinti ad relatively ordered steady-tate of glahim výhen the added ADP a converted to ATP through oddative phorylarylatka respiraton s ccn to slow down markedly Thas change la rclockty of their trosport a accompanied by a reduction in cytochrome and the are now wodt bom Stat 4 the querent state. This watch characterian by the presence of substrate and by the abuence of phosphate aceepedia There acrobic metabolic state de mitochondria are well kormwn from the la comtitate Do DCW Inforetan lorrrumper most of Chance and I am to point out that the metabolic sulas described for other mit www.ralp tere are equally relevant to beurt Broot at grefy about the State 31 ftrenution, bom ever be the reply decrease in tortility in the world ended (1 410 now oly) trace the b due to the meleg athuman It sagribcant to note that the marcoma eman in the shrashur interye state only look u the ADP added 's betos phombarybalo heroes ·520 Andals New York Academy of Sciences the citent of the light absorption charge caused by selling is very much larger than the specific change observed rith Stochrome 6 and the bigbt uborption change would modeed have been very much larger than that shorn if it were allowed to continue a ithout interuption Collowing tbis, & mcond ton of 250 - ADP u made, awong a State 4 to 3 trungtron. The follon ing events dotingunh thu change (1) taputoa increused by factor tenfold grater than that obocrved in State 4 (2) thos Increased rate of electron transport repults in an oodation in the stady hate of cytochrome b and (3) a shrinking of the parconoma taka place Vitochondnal shrinking takes place at Ante Hoser than that of cytochrorde doudutron It can be seen ibat tbe mrcomnes did not shitak to their former due tha was only because the small amount of ADP added nu converted to ATP in approumately 45 seconds that ., borger addition of ADP moald have resulted in the complete reveral of malling us shown by other per menti When the intimitochondrial ADP concentration decraks due to pboxphorylation, selling begins yun, and the other changes chancteristic of the State 3 to 4 trositron are clearly sen. A third addition of ADP accompanied by Hailar changes The two complete cycles of ADP addition are accompanxed by ADPO nattos of 3 2 and 31 and resptratory stimulations greater than 10 tima Theme nition are easily calculated from the amount of is utilized during Sute 3 cod the amount of ADP added these results are in greement with the high phos phorylation efficacy of heart Artosoma reported art by Valey and Plaut and by other Altbougb ATT berng accumulated daring the accouve perlada od pbauphorylation shown in the esperiment, the rate of swelling of the Artolama i umffected Indeed, the addition of ATP itself appaan ineffecthe in controlling mitochondrial wre encept by virtue of its abuity to formu ADP Th, the addition of Big rous to wrcoromos incubated in the presence of ATT results in A Trux activity and a consequent reveral of selling Other cıperiments thus far have indkated that ractions tbat TVC to decrease the intrimitochondrul ADP concentration cultun melling This, orduable substrate in thate swelling, and the krpetlas of the rolling recimo ere found to be related directly to the rate of respirttion Conversely reac thons that increase the intromitobondral ADP concentration reverxen dling some cumpla of these reactions are imaoblogsmbibition of electron trans part, ATPase activity und erhaustion of oudisable substrata In conclusion it be beca found that the intriumitochoodrul ADP nontntion bs involved in structul and functional changes in heart surtomode, u lodged by the amultaneous aurement of mitochoodrial wc reputa tion, opdative phoxpborylation, and the Andy state of the reptratory en LYTch, us cremplied by cytochrome These finding date that ADP may play a primary role in the control of these processo Rdos L RULADI, J 1953 Hetv Pol Pharmacol Acta 11 12 2 REITL4,) 1953 Ich hy wol Pharmacol Act 11: 157 3 of OLAN, OŁAG SIND 198 Brochen J H1 569

·521
Pacher Structural and Functional Changes
IMCA, L Foro URED TIS 1956 Mladro J 1754
SMIL, RF LILA LOTTORE 1956 Botimet Blond Acts 34
6 LAO A&H MAU 1952 ) Bol Chen 16215
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1950
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1986
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0 CUBEL TAT 198 Bucke J. 193
1 SLALIY G FUGITZATT 1953 J Chem to $1 191

·520 Annals Nem York Academy of Sciences the extent of the light aboorption change caused by selling N very much larger than the specific charte observed with ytochromed and the light absorption change would ideed have been very much larger than that shorn If it were allowed to continue without interuption Following thos, a second edition of 250 V ADP * as made, courg a State 4 to 3 vandton The follom og cyents dolinguish thus bange (1) respination to increased by a factor tenfoki grater than that observed in State+ (2) thos Increased mate of electron transport results in an oddation in the seady-state of cytochrome and (3) 2 shnoLing of the Barcodon takes place Vitochondrial shnnking tabo pla ata rate douter than that of cytochrome doddaton. It can be seen that the an doma did not shrink to their former die thus mus only because the mall amount of ADP added us converted to ATP in approumately 45 monde that B, a larger addition of ADP would bave rosulted in the complete reversal of maling u shorn by other open ments. When the intramitochoodral ADP concentration decretos due to phosphorylation, swelling begins again and the other changer cbarcterfall of the State 3 to 4 tngton Are clearly sen A third additional ADP accompanied by ardilar changes The two complete cycles of ADP addition are accompanied by ADP/O ratios of 3 2 and 3 1 and respiratory simulations greater than 10 time There ratios arc cauly aak ulated from the amount of utilized during Suate 3 and the amount of ADP added these rulu ure in ugreement with the high phor ) phorylation officiency of heart harcosomas reported earlier by Valey and laut and by others Although ATP being accumulated dunng the forceTVE periods of phoxphorylation shown in the caperiment the nate of nudling of the Arcoroma is unaffected Indeed, the addition of ATP itself appears ineflectne in controlling mitochondral size, colept by virtue of its ability to form ADP Thus, the addition of Vig som to mrtosomico incubated in the presence of ATT results in Allu activity and a consequent reversal of muelling Other crperiments thus for have indocated that reactions that serve to decrease the intrimitochondral ADP concentration result in swelling Thus, axdiabłe pubertes unitate mding, and the Lunetas of the medling rau tome are found to be related directly to the rate of respotion Conversely rac toos that Incrause the intramito bondral ADP concentration reverxe relung come crumple of these reactions are ne crobroms, inbibition of chet tron tren port, ATThake utrvity and cubeusuion of didiable wbuntes In conclusion it has been found that the Intramitochondrul ADP 00cent truons involved in struktural and functional change in heart arcumch u fudged by the simultaneous manirement of mitochondrial are respira tion, oudative phosphorylatban, and the Hendy rate of the repintory en Tymes, as eteroplified by cytochrome b These findings indicate that ADP may play a pruary role la the control of these processes Reformes 1 RAATLIDT J 1953 Het Idharmaco Acte 11:14 2 Runom, ) 1959 Hchy Phyo Ibarco Acta 11 157 s lucturLAT, NO&AGS 1933 Biochen I Hiko ·Podolsky Thermodynamics and Molecular a lechonum 523 R ܗܐ ܫ Recovery proch, Lolt al proces TOULE 1 Ractua chon far cycle of Dwalar actinty The release of cbemal energy reflected in work and heat production Therefore we turn to the clegant erpenments of A V Hill, who sorted out these serpy changet. We shall consider mucles of about normal body length and me that shortening small that chanpes in length-depeodent pann ten, such as tetanic tendon, an be eglected This means that even though the muscle shorters, the leagth always remain in the region of the broad must mum of the tension-length curvo (Rumky und Street, 1940) Froun 2, from Hillclum 1938 paper (Hill, 1938) sboms how the tenslon developed by I musclo depends on the speed fco traction Th odlotos the shortening velocity in wolont rontration, and the abacus b the loud, or ance the velocity 3 constant, it is the force cetted by the muscle. The intercept on the abecim the tetank terigon, The mooth curve the well-known, hyperbolxe free velocity relation, + .) (+ 5) K+ ) where a add 6 are characteraix costants of the muscle. The nato of doing work, and the produrt of the force und the rebooty that , the product of the abacte Lod the orderate of TOUZE 2. This product Voubor both when there Holaad end ben tbe load exceeds the force the Muscl an develog so that the muscle i prevented from sbortening There * un ntaneduto sborteolog rute that parmio the rule of dort work. Clearly th rato at which precbanical energy lare the macho la controlled by the speed of shortening ·524 Annals New York Aademy of Sciences itoaty of shortening on loc 20 30 0 50 Lood FRUL 2 Relation between ibe rooats of shortent, and the mecbunaal land in Lotuc coatncto Reproduced by pero non from Process of the Royal Socia'y of Law (Hill, 1998) Energy also leaves the system u heat. FIOULE 3 shots that the speed of shortening abo controls thls procen. In ach case total beat output from tetanxcally alonulated muscle b abonn su function of time before dunng, and after abortening in the set of operiments m TIOURI 36 the muscle relausod 1.2 mec after the start of tetanic stimulation and allowed to shortca vuous distances under constant load Curve A represents the total heat from an bometnc contruction curvo B C and D represent the heat for at crudng amounts of shortening During shortening the nate of heat produk tlon increuses relative to the ntc for sometre contnction and the total ein hat is proportional to the upount of shortening When the relocity returns to zero (final sections of curva B C and D) the hat flur u cmentally the more in the shortened state u n the corresponding sometnc Hate The beat from the ame muscle shortening & Constant distance under Increang loads is shown in curven J HG and Frospectively A TOURE 36 The muscle su relaxed at 1 2 rc the end of aborteolog biodlated by AITOR , Curve E represents an isometric contraction. Since, at the end of abortening curves J H, G and Fire Jl dagplaced from the women contruction by a comtant amount the atn bat aroclated with shartcolag does not depeod on the velocity of contraction, but only on the distance shortened Thu means that - * -u the role of hat production, selcar function of the speed of shortcourg -0-sto ·Podolsh) Thermodynamics and Violecular Blechanism 525 (a) 1 1 1 Time marks 0.2 sec. 0 F E 1 Time marto O Zsoc Froce 3 Toul bent productions, -04 fraction of tror Tetawally whated pecka OC Scoton for the perpetual coachnops Reproduced by pero from Proceeding at !! Roger Sandy London (AA, 1937 The constant where to the nated beat production in munc contnction • s dimensionally and aumencally the sim utbee in the force codity reba tom (EQUATION 1) In the follor in ENCADOU 1 and 3 w ill be referred to u the empinaal (cseres relations of Hill The cperiments indicated in and 34 an be summanied by aymg that hant flur from Rimulated muscle depends on the speed of shortening A contnbation proportional to the speed of shorten og b rupermoposed on the bent of activaton th bawl nt for Amuhted muscle constrmed to remata at constant length The certy manifested on ork and beat derno ultimately from the chemical cnety relaxed un reaction A Th modulion of the energy flur by the speed of shortening imples that the nate of the dras me reactions also Imhed to the De parameter To work this out na tally begon nh tbe first bo of therproduk ET -- bere all sth fange in enthalpy bin the entent of reactboo A In E AT to the change in cathalpy for un tentent of traction, and eka the ack dooc by the Aston DCT * of the randon work, PAT Different τουΑΤΟ + 1b get to the ETI 159 --- ·526 Annals New York Academy of Sciences Loco A con generally be treated us a constant quantity The tons on the right hand side of DQUATTON 3 can be obtuned from the empirlal relations of Hil. Combining EQUATIOX3 1 2 und 3 with 5 ott Echo 5+, where to - WAN the reaction rate when : (ind consequently w) is pero uod 8-1 + b + c)/ 1 collection of constant parameter of the muscle. Thus the chanical reaction rate & is a hyperbolk function of the mechanical raction rate, EQUATION 6 an be evaluated oumencally by using the empirlaal obarva tko (HII, 1939) 4 - ab (7) and € (8) Tben B is equal to 6 and the marnum contnction velocity : (EQUATION 1) equal to With these relations EQUATION 6 tala the ample form 1 + 241/C (9) 1+ te/co When -. that is when the contnction is unloaded the chemical reaction rate, at laut in frog artorius Muck, IDCTEUs five uma relative to the nate for momctrle contraction. This relation is plotted ut olid line in FIORE + The aburton is the relative feed the ordinate, on the left, by the relative renc tloo rate. The dotted line is de/ds the extent of reaction for a given amount of abortening, and i akulated by dividing the chemical reaction rate by ube mechanical action rate. lle see that for a given kount of shortening conaderably more chenkal energy must be made available it the Duncle mora Howly rather than rapidly This mains that the driving racco AB Dot goured to the displacement of the muscle like a nick and pinion device Rather the mechanochemical coupling bs more like a fuld drive that permiu slippage between input and outpat shafts Kindios of Vlada Sye What chemlal sygems have these propertla? For cluas we turn to OKTOkope studies of contracting strated muscle Photographs made in Rucky und Niedergete (1954) with an nterference microscopo ilonte the changing stradon patten u aliying murke contracts boloocally (roz; 5) Short cuy from top to bottom tako pb e almost entirely in the light I bund, the indth of the dark + bands remaining content Thus thom na mort drumatkally in the etperiments of Ilurley and Taylor (1955) in bkb 100- tructoo confined to i angle arcomere ruimulated directly with a micropt ·Podolsky Thermodynamics and Molecular Mechanism 527 erilo 250[dx 20 2 2 02 04 06 08 10 Ta 4 m run to reaction fonction d the roletne loot of contraction, - Corresponding metal for shabang nodd are plotted ahdo morta for frodeckans grond are plotted to a Porb Dadete Rre ak to code ith the booth (-02 -10 (dowed arda Del com relation bet en tbe atat action for at Koutrako, A/, die vonaty o Conction pute (TETRI 6) To pucture on the right shows the popate Det I the 6bct before Himulation As shown on the idy, when current speed throogh the pipette, depolarning the membrane, the I band decreant to Ilth antu It remishes, drax mg together adjacent A bepes Studies at higher magnitation with the Hectron MITO cope port to a filament d aru ture of the muscle Abril thoxa dumamiatkally in INTRE 7 The main point t not that there are lo Lunds of Glament hose peculer dosribution as the to the charteristic anatom The t band duct filaments that appear ibrk in the electron miokape and ure bought by some to be made of my own The food stotlements, thunder the the fint and poubly made of setin, cated from the Z llore through the / baod mi

·528
Annals New York Academy of Sciences
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·529 Podolsky Thermodynamics and Alolecular Blechanim ܨܝ MU MI 1 From 6 Shortening nogle / band otaced by dopoldne odlarea with micropuppete it apper edo al bor) oght, belart dhepokantation kalt, alter depobertation Reprodord by neobon from Jurnal of Phnogy (Ilock, und Taylor 1953) the A band, there interdigitating with the thick filaments. The thick fila ments are definitely discontinuou the thin filaments appear to be continuous It boogh tbey are much thinner near the middle of th d band, which is proud bly the rouson for the loner inde ordrection of the I zone. Both deetroD-Microscope studies of fired preparation und light-macroecope observation of living muscle suggest that during shortening there is relative motion of thin and thick filament In pund by these finding, A. F Hudley (1957) noted that of complementary sites kere located n the thin and thick filaments repectively and interaction buta cen these stes depended on the relative position the Lipetics of interaction wul depend on the speed with which the filaments pove pest ach ther Furthermore, if the interaction between these moving atas uere chemxcally hnhed with the driving reaction, the could cpuin bow the peed of shortening mfluence the nat t bich energy s mopplied for the contraction. The curentul fatures of tho da art bor m TIOR 8 The thrch and thin f mterdigitating flament linea reprexat the thick and thin vet urt on the thick filaments and stes i are on the thin filament I ute are complementary that some chemical nteraction can occur boe then tbem The interaction probability per umt time K) bormed to This schematured by the dotted, depend on the relative Porison of the ates bell shaped curre Suppose the thin blament moves put the thich one then play Sita T Tbe T und S-filoment for 1 Dund Decker Reprobed try parconio Program B-MTK (ks 1957) ·530 Annals New Work Academy of Saences Tuto Tuito Tute K(X) taito Four & Thad and thin semonts of muscle fibril See tal for description For et 112 i ste approaches 1 T ste the probability of interaction dur untu the ata tre directly opponte each other As the motion continues, the motor action probability decreases, finally reaching zero when the distance creede the nudlus of the bell. For a givea amount of abortening, there will be more Interaction the lower the relative velocity nince the extent of interaction u proportional to the ulme available for interaction As shown in FOULE 4 thus is what happens, qualitatively ut last, in Irving munck. We do not know the nature of the T+ interuition Tortunately thu doa Dot block further amploration of the model because, in order to punh ahead, need to know only the stato trcal nature of the interaction. We shall assun that in the course of a trangt of u ute pesti Tute the micractxon event (1) alber doas por does not happen, and (2) aan happen only once Sample, oppose is a binding ute for adenosine tnpbosphate (ATP) and T Joan ATPuse nte Then, us the utos nove past each other the ATP either does or does not get out. The probability for interaction (splitting of ATP) grater the Hon er the speed, but the raction can happen only oncc far auch molecule. If thus reaction u the nate-limiting rep in the chain of coupled reactions comprasing the initial process (TOULE 1) it can control the rate of the reaction sequence In this are the THintonction nate can be equated to del To darve tbe kmetos of the T+ interaction proces, considera pur of atos moving past ach other with relative velocity » Suppose the displacemeat of I relative to Tbs Then the probability that interaction will bave occurred already b. The probability ## that I and I will interact in the next element of time as the product of (1 - *) the probability that they have not yet interacted and the a priori probability factor for Interuction or 1x = (1 - *) W (10) Since the datance tarcted in the demont of time to - (1-) It/ (11) Intention across the entire transit gives the probability for Interaction in a complete tnnit at velocity * (12) bare- [ the ura under the bell-shaped probability curve. The ·Folding mochanism are med to be fi ad rcn to tbe the filamcats dire the PRC atbo micteos from a chance to the elustiilty of the As opposed t the Jding model bont the rath und Tato non depends on the poin bong the filent US 10 Mechanal pring the reth velocity Intan Enarty she Podolsky Thermodynamics and Molo ular yar A I А tron top and bottom, CO Ular coatraction result o intulivdy metafyon bon tbe qeeda mfin tc, tbe expomenitih anity and the probability for intencboo, * vanbben. when u bo zero the ponential vapabes todo unity. We o dotace that the perkubur sehen he wants the probabality curve does not matter Ia practple the curve could have had nurymmetrical shape instead of the symmetnaal one chosen for Ilhairathon Before thus analyus ad be pplied to contacting muscle the distribuidora de utes and of relative clox ty along the tu Lods filaments must be deckthe 1 shall come that the sta ut dounbated with muform bear derady along auch wet of filaments Each Tt interaction augumed to be independent previous rotantin The relative velocity of atos look the two sets of filaments will depend on th relative motion of the filaments during contraction Corent models for cootruction can generally be damed us ather sliding or folding modeta (rovar 9) I both modelo contraction is accompanied by shortenbar of the word the i band wodth remaining constant, on esteement with the microscopecal In th dadung model the cootraction force urbanes from interxion cvidence The relative velocity the eam abort the between thick and then filaments In the folding model the end of the thlo arrots YE eod of th thin filameet the relatint iclocity ocho If the follo entire region of overlap The for mcats 1 ment, reaching ma waum Dar the 1 bard.

·532 Annals New York lademy of Sacaces Do these differences in the contraction moechanan land to corresponding differences in the dependence of the intenction rate of the bol murcle on the speed of shortenung? The ansa er depends on how much se know about the Entertction eta If we try to bt the predictions of the Hiding and folding mechantara to the thetibodynamk comequence of Hill's energy relations, nc an do so rematably nel mith both modela (TODE 4) The circles abow the dependence of the T+ intenction nate on the muscle contraction rate thing a Aliding modd the cross mere akubated from a folding modd (APPENDIX) These and be compared with the old curre derved from esperiment The modid points a ere med for fitting Here that both modela can be adrated to fall wdl sithin crpermenul uncertainty although I a choice must be made on matheutical grounds, the folding mode provides a slightly better Apprunmethod to the hyperbola than doa the widing model To make the cakulation agree with the hyperbola at relathe clociues of 0.2 and 10 different valua must be asumed for the dumber of Tutos per unit volume and the dotance between I rtes (APPENDIX) However unce we have Do independent rules for these qonutla, tha cundot be used to differentate between the two ontruction mechanKT: We must conclode that the chan- Ical krpeta denred from Hulls energy relations can be accommodated by hark model with equal facility ular leven The contraddle Blechaxum We now consider the interaction between the moving filaments on the molec In the widing model the interaction between filaments generate the contracule force (Hurley 195 ) and also regulate the fun of chemkal chergy The ate on the thick filament u placed on an eallating "ude prece huch can attach itself to a complementary ute on the thin filament If the attachment is made when the ancillating nte o to the nght of ius equilibrum porllon, it will tend to pull the thin filament to the left Corvetkely if the connection occun when the ode preces on the other ode of its equilibrim position, it w tend to pull the thje filament in the opposite direction Tret unidirectional (ahortening tomaron it must be usumed that the probability of attachment and detachment n symmetrial with respect to the equlibnom position of the oscillatin etc Hurley sboned that of the approponte To metrial interaction probability functions were postulated, the temonould depend on the weed of shortening 10 the way described by Hel s forte velocity relaton (CQUARIO 1) The folding model hos ett requro no exch resunction Agus, for o ample, that the contractul face in the thio glament Kerns from n unbalanc in the clectrostatic and catropic forces in a fleoblo polymer Roseman and Kutwood (1948) aod llorales and Bolts (1952) made this useampto in dor CIBAN related torlek The argument prevented Legna sith the wharged filament (ruta 10.) The cock of the flamentul teod ton and the pollon that maumires the nuralıt Of configurations the filament consume Thн tendency the entropk fonte allan the word of the filament together Hox Tatlo model, bonne coonts for the bouto tune, ·Podolski Thermodynamic and Jlolecular Vechanum (a) Uncharged (b) Chorged fibor fiber 533 Tenson ə + 20 + t 2 (c) (d) Foon 10 The electrostatic-europie mecburst forget teosoo e particle CUTY on out to barre (a rocharged flame folded by metropic force b poutney cbarred Glaspent a uklon kehitect for at balanced by the Latic foret () (d) and Depu ch deed putte bound to postnet ferred Silinment Wet om dut u Teproe din cloped by kmet proportion to en Marution o carr ever of burger reduenbuted along the best the entropa for mulbe opposed by an der troudalk force tending me the duten betreun charges The cquibrum lengib urhes a balance belu een the entropy for itching to aborted the tilement and the electrostate forte iendmy to preteb 1 (nom 106) If dkpokana on the muscle membrane mala harges that an exit the filament the filament al tendi the clectrotutk force u bo longer balance the entropoc force (TMULE 10c) Suppose that, as the flament totens, the attached charge can be detarbed by tetection with passing atas For comple, intume that the neqat ve churra ure amed by ATP nokcules that stick to the i pta on the thon Gamcat As abone the intentoo »ith the Tot on the bad filament (not goud 10 TRUIE 10) CRO be bydrolysa followed by desorpion of tbe produkts Then, fter each interactwo bere null be a fnite time of the abble negative shortct once ·534 Annals Net York Academy of Sciences P/A 10 03 O ROH ME Figur 11 Tbe relative ton, 1/4 functbool the rate a T4 altricting tbc latter is beta cqmated to the nicol cbemacal reaction totne toon. 6-mat al cepat reaction ben the contacto coaty sero, t - rute of characl recto ben tbe contraction daty num nert ATP Hicks on, when the inte vacunt In this way the average number of ATP molecular on the thin filament at any time mul decek u the interaction nato increase the simple mechannm relatos tendon to the interaction rate uoce, m the first apprormation, the tension de cloped by the filament nl be proportional to the mount of Dutralization of power charge by negative charge adaorbed on the intes (FIGURE 10c d nod c) Thu the relation between relative tension, and reaction nte, Eshon D 10 7701722 11 folloni quite naturally from the dectronlanc-entropa machanna of tension development. The tension will be mutuum when the chemical reaction rate is miomum, for then the poutre charge on the filument fuffar the gtatest mount of charge Deutralwation by adsorbed nequinc charge. As the reaction rate inta, docteung the average aroount of peutnalization the tension decrca Stated algebrawally & - 60 e- (A -) (13) A where 6 und are the reaction nates for koppetc) and unloaded oor tons, respectively The modd por complete and can cuptain quantitatirely the dooty dependence of the mechanical and thermal Merty fluo ja Intact Living The argument raus on the sumption that the limiting factor in & the rate at which chemial certy boude inilable to the contractor tbpoe the nte of action of rubutacas coustrained to stay in toatht potlog, that is the rate of Tintmction Since the Linctio for mieruction of this type xert somn to be homeomorphk with a byperbolic relation be The bad nooze 11 rokdo that feln-5 ufrog artono prock muncke ·Podolsky Thermodynamics and Molecular Mechanam 535 Inen und the peed of shortening. - (none 4 und APTTYDX) the (anc thoa (t) can be approsted by MUATION 6 The fanction () Wkea from the relation between cbarpal action rate od odon, boherent la the electrostat-cotropk mebanen for developing the contractile Corse (DQUATTO $3) The characteristic of the cock can then be obtained u fations The des force-relocity relation (CQUATIOY 1) reell from climla tion of beti eco CQuinov band 13 od the Woulfillon Uning the force-relocity relation we aan akulate the nie of doing cake a function of the speed of shortenint Then the beat production, can be derived from the art and thermodynamo (KAUATDOS) und the kinetia of the T+ teraction proven (QUATTOY 6) I would li timphatite the fact that my audio s not welded to the dectrostatkeaurops mechas won for development of the comirectlhe forceIt na honen omply to allostrate a dars of chemism in brch the contractile force varia Imounty with the amount of mooc molteule bound to the then filament, and in which the bound molecules can be borwwbed ofta la paredne ata on the thick barnent The remarkable thing about the kind al method that the characteristic of the living Umuc are derived fron qute general romdetalkom The orect equatro Bow from the trurtur rather than the details of the model are of course a mamber of dsturbing love ads that need lying, but I feel that much an derommodatne model derives to be an ibo that of the opport There txon WITADEX Tu Doppia o THE TITATION RATY V rochuv nociNY KOL Surara w Taarvo Voorus or CORATION Sub 1 studo beatp tutos on the then the V-bar Tato/a onde - Tiberioare a oroda LA T7 odszed contraction o noso edital coro, the per 1 dera u Uprethtu te punt the code of the the blue Mount Aral nth to hand dhe Used the contended by boy bebe DW (Hockey und Havn, 1944) portioon to be loggebodabode of decreto legtes Homme and there Do cerrado con bot for whates the based on the internet and Perry 1991) second potato cotove be both us ( wy model) or od of my for the development to codecte, che the ATP oral contracte con el extraction of brand, store mely the loc tots d pastrolar The aparental moel tro o char webes the TT populo Rinot ticted flady od Home 1094) buk bobots that Luted contracte pou Addition of ATT heils au Datbe Theor.cored Contand Part 1 The yine o coure a la recepata tayo at de caput i the future rotrop methoden god bentChath the pour mon further aporta study ·1 536 Annals Nery York Aademy of Sciences - probability Thutmcte in complete that nabo of the distance of site from the red end to the length of the follog ropo (m tbe fokting model) - laptha mercompet - roogty of Ixturdain to Tita - velodty of contract of the ads of the mode (apromed in mouche kosth/mc) - na mder the probability care for pron Tintention STRA waid I thoshding model be rebaty auto on the thin filuncat relati Tdtes on the thic, float s thenealoog the length of the flament Torreloaty related t the nte o sbarteoung alte mtzt mack, by the apartman - (15) bert is the nrankere lath If the distance between / utaslach Tat 111 be pet: Kotod with Axista/uat time Therefore the iteraction nut /cc of puce nu bat the number of Tita/a dmuck, und u the probability of Te+ interneton LA tot treatine relaty Uung the function () from po rio 12, the amboo Ti bateractions/cc o mock/umt tune u - (1 - 1) (16) Caabution of LOCATTOO 15 th 16 gives thont of chemical recto us function of thx speed of bortaing of the code of the macho гена 1 tbe fakliy modd the reloaty di atas relative t the T atas 10 bo od ut th iced cod of the thin filaments und bo t the end the folbing renou clonout the I band, bort, us in the abding model, tho sbort roup Poaty d the Murcomat 2 Therdore, if the nato o the data col pre inte from the feed cod to the length of the folder reron, urem velocity ben A Tate at the correspochne part of the thxc filent flufer /l trants, bare i utbe costante beta con i mla Thordort the ronction nat /ce of monkey N- (17) Carrying out the hotaratna, N- (18 2 (- r+"[ *+ **(--))) TE) w bere (-) the opportual memul The function Yout from cobertation of QO por 15 with 18 For the Holyfolay meds to be hypertel hoe land for 16 ww porno ratan. It has beo box that the nat o the driving rector for matnction by me boho fanction of the shortening speed (IQCADO ) T compent to function it tho Lip tra of the Lung und foking bodel, ale vert foncn for the qunutos luoda to pak FOO to 16 and 18 me to the hypobolant-02 and A - 10 Doctent la dulu mated for and model (V/ TV-04 - 20 In loy chected woody ert aboutal according to rotanov 16 sod 18 d plotted n anda dan petirely a nav 4 Roeroues Cors, ISIMU 1955 Some OLET Iloos on the haluation d 01, etu otropomun in the blat Doblo Brocheo ] R12 1939 I be bat d dortmunt Add the dync COLLAU O anche Proc Ros Soc London H: 136 EL AV 1999 Tự h a d d fg sẽ Proc Ror Sre Landon US 144 JIAL AY 1951 Tberpody papladnock Veturt. 187: 377 ܙ ܕ ܝܫIfn

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·VUSCLE PROTEINS AND ENERGY UTILIZATION I Gergelyf Carine Blancois Lord Lady Departakiw Vuokatuoti Gourd Bespect and llanand Wadical School, Boston, lass I wonder if tbe difficulty that one faces in trying to correlate the structure and onrymutk uctivity of muscle protams with muscle function and with the utluation of energy in muscle us sufficiently realued Ordinanly the enty mologat bes a fauly trughtforward tesk He begins with the knowledge that certain organ or in le performas e certain btox hetoxal function he then proceeds to prepare sluca, breu, and entnu ts from the trade and in solating i miten heb gunded by the chemkal retition he has in mind Honever the muscle or heart blochemat or phyuologata in a much more difficult posuion, uh u interested in the entire phynologal or bropbyncal prooems of contraction. He soon encounter a form of the complementarity principle The principle 3 a fundamental one of modern phy, and it Keto certain lim ts to our inquinas and to our knowledge A stated by Noel Bobr Evidence obtained under different experimental conditions cannot be comprehended within a single pxture but must be regarded as complementary in the sense that only the totality of the pbenomena erhout the pooble information about the objects Thus the behavior of light can be described depending on the crperimental unungements, ether as I have or as a strenm of particles. As another example, it is impossible to measure at the same time month very high accuracy both the portion and the velo ty of in electron Either however can be determined wartety Th Fort of complementanty may be operating when one yoxdies manche One may obtain the type of Dformation that was produced by the briliant work of Hill and ho schoon who approached the muscle with the tools of the lassmal physologi t including the most intricate measurements of thermal, electrical and mechanical phenom oru, If bowever biocheroust approaches the muscle be will turut tear it apart and with luck, get some brochenkal information hom cvet i will be extremely difficult to my anything about the place of the hemal phenomena in the context of contraction In the past by the use of these two approuche much dctal Information ba been accumulated by the pbyoologists on the one band and by the biocheraists on the other However tweem to me that today's most urgent tasks to attempt to achxeve a syaibew betreen muscle physiology oltrastructure, and brochementry lle can only hope that the powsible, and we can at last parruc our inquine to the limit vet by prisciple similar to that of complementarity The art reported la this paper upported by mnt trom the National Hart Iosu tote, Poble Health Sers ice, Betbed, Vd the Moralar Dystrophy Ascono al Amena, Inc, Ko loth NY the LP lost sledial Roab food, lo lock V3 the vocboxetu Hart Associatba, Boston, lo Thepot en chans rekes the object, bat merity duvao o kome problems of each broccouT) thet en al perthouer tolereet the other bon march, tout pou to be particularly dcult to pok The reader det en faber one tion til ordered to LDY aed kon met collatok Esbedinator of the leau llart Arocuon, Inc Nor York NY ·Gergely Energ Uuliation 539 It should be poted that the panciple of complemediarity in its proper that to cranium mechanical sove ray aho be plagaing The concepts old classical cherdhiry and phyæna may soon be inedespate ho describing no in trazte system auch u muscle and we shall have to turn to the proper use of quuntum mechadas a we hope to achieve a vuld description tic om tremendous debt to Albert Saat Gyorgy? who has been a champion of this kan for the last thaty jan Ia any attempt to bridge the gap betalen physioloos und Wocheerlantry In the axe of mawacle it mupparent that the fint problema liat la deciding (1) which proteins and cozyme thicons that can be obtained from a munkcie ertnct bare a role in contact on proper (2) bich are pombte artifacts and (3) which function Derdy a torpecific suppliers of mergy In the form of high energy phosphatompound I think the worting out of those proteine and crymas that fall in the third cast a relanvely almpte loc we hoow much about the procera resporable for the generation of energy both in the present and 10 the abecoce of tyen tbex procerxes recerre conuderable attention clxhere in thr monograph Therefore ben we upal about proteins pocetoed with muscle contact me think in terms of those protina that on into their own fter the proteina that supply energy are done their yob Recorded AREN'S ***.pcla resoch The last few yan bave brought uremendous advance la thu field then doet veral factor Feme, there was the discovery of certain murte model, that w systems that stand balluny been eeu the whole muscle that was the object of clerocal phypology and the pmtan and enzyme propunation studed in th brochanical Laboratory a result of Albert Sent-Gyorgy dre overy it skoon that one can einer mowle with 50 per cent glyrcrol od Hort it for almost indefinn period of time a deep freere, and Aul obtain ontruction When tbv fiber having been removed from the trees and mashed out with approprie welt mohu thons, u brought together Ruh adenoude urbomphate (ATP) obe obtain rood contraction Tbe analyses of the contrats bas bero cursed out by mnoon groupe Weber und bs mbool op Germany have done a purtxularly thoroogt job in sbon my that many d the properties of the contncion of the glycerinated fibet te bented .th tbox obecned to living muhe Ono phenomenon that partxularly top 100 x the so-called pbenomenon of borter qarch release If a gwulated mak ddenly roared to Lagth, the tenson will drop conoderably but after this adders drop there *ul bc e non son 20 a new equilibrium fere come pandong to the ser length The aume phenomenon CD be obarved con glycermated muscle fiber Thiero are other unties, and treatable to my that the greenasted manche aber au be canadered fatty good Imh bets een the abole muck and the oluhle yuton abo occurral Alany ad nos in our knowledge of the muscle protein to Nek mus le proteina bure been decorred, and fragmente a thoxe Lnown for It sulodeat that before ane omparsons between shole wel od the procesos entraded from the one most hoow much more about these protein, bel gructure, and their lope Exco Lurbed kone time be ·540 Annals Not York Academy of Sdences toc The third are in blch great adyama have been made # that of the foe structure of muscle. The important fatura of this work are the u tempts to determloe the locallation in the muscle fiber of proteins obtained IN FT The fourth are in a blach interesting developments have occurred in the lut few yan ko n the study of chemical changes in muscle during contraction Hark of thha type has been done in various laboratories, including those of T TH Jlommorts, Unreruty of California Medial School Los Angeles, Calf Carbons and my oto un collaboration with A Cup of The Rockefeller Institute, New York, II These studies bave rassed interesting and per plering questions, used nature, so for bas refused to give a direct answer to the question that chemal changes occur in e angle twitch *** I now return to the dmpler things, which actually turn out to be Dot mo simple that A, to the protams that an be studked in solation The term myou has been in use for nearly one bundred youn, baring been introduced In 1859 by Kuhne' although its original manig was different from the xnge in which it later rame to be used Until the docovania of Albert Szent Gyorgy and Straubte in the arly forties, mayoria denoted the substance or substances that could be erincted from ground mucle at a fairly bigh hona Hrength, about 05 II but were moluble at lore lonic trength In 1930 von Vurult and Edsall" showed that axh an attract possesses very strong double refraction of flow indicating the presenxc of elongated, rather large, molecules In the meantime, the lactic acid theory of muxl contraction had been dir carded and the phosphate-contaming compounds that today are comoonplace to biochemisu ane to the forefront of interest During the late thirties and carly fortias several group were able to sbow that myosin, the vz.com etnct obtained from muscle raudue, bows some remarkable rauctions with ATP It wu discovered thet, on the one band ATP ha profound effat on the physxochemkal properties of myova The double refrac Lon of for disappears, or consderably decreases, on addition of ATP od threada prepared by squrting myosin into water contracted mith ATP On the other hand, ATP tech lost a phowphate group when it *as brought together with this muscle extract. There discos tria) ure associated with cedham a al Engel hardt and Ljubumova," and Spent Gyorgyl and bo co-workers Thus it was apparent that ATP can affect the mechanical and physio chembual properties of the muscle proteins, and that the proteins, or the proteto u it au thought to those days can in turn ect ensaymatically on ATP The tempting thought cross that there must purely be the reactions that underlic muscle contraction. The work of Lundguard had rudy ubown that mucle contraction by somehow Inked to the break down of orpor phosphate compounds Hyer-B and reconstwa low UNIR It was wou found that the Altcore re-wanted myoan-B by Szent- yorpi and his odle gues' is not the only interesting vulsta xc obtamable from the most resdue it u poodle Ao loportat de cover lang os prowon ameck In Dar Du logo are the prepunbon of the paper ·The follows are Gergely Energy Utilization 341 to obtain a protein that had the ability to HATT but the protein sboned mchange under the nflent i ITP They saw trollin bt am Inn In tal name mimin il man, milline myrir, or simply mytun Still styros in un nixostra id from mache unit anteact with man to mes frondom i that I try smlar the old grytindetin Iball be in two forma gkbular forms and abbrain form. Unok the Influence of salt the globulari dormed into the fibeak form It is the latter ibat atera ! *uh man and form het sy be called seronstituted actortyusin dyoum-B the old myouin has been conudeted in general to be acto mjoen Indeed at behaves sery moch Rex combmation of actm and forain ali barangh watu secently there has bot been much direct pmor that it actually Come of myogn and actin Some work stim good circumstantial evi deacent i lavor of the new but it on important to with this guestion definitely One studies the combination of mynds and actin because one believes that thx umpler in the same * myounB that one caructs fronti make and that a belseved t the contract le groter One must be surt A Weber has conducted one experiment that there to gp in the bain that, to my mind, make this poutro mong Recently however oming to the work of on Hoppeld d nd of Komins e al the problem of the klenuty of actomy own and youn-B has become other crurnal experimental dats obtained is my laboratory by Startonas Is appeared that the amptent eppach would be t attempt the moolathon from myo-B of a subrave banor the properties of actin Bartonon! wucceded in dosag tha He prepared my own-B frented 11 with acetanke, isk then extracted the dry acetone ponuder Ib dostallet sater in tbe sac nay e doat in the propanin or lame luctm except that sho latter en tnton, one treats the muck redue well with metoo cheur wolution of bow wouly that, oa adlawon of ult phoned an trouve la Voiconty the bas been known to be property of Act Furthermore, thu muabetarxe ben added to Dran, creand the yok oly and the intente ett resand by ATP Jforever the of the scattered with both beppes proseso sabied from myomo-B krungty metrused the 47 Pase activity myoon at low wax arength in tbe prosess of light (TANU 1) Therefore m all these respects the solated compound beloved ery mub luk remune 1 tbea turned to one of the most appear for thethods in buckyy umunology been brilliantly put to use by Holtmer and Inmunochemical met bort he Blansha!? * B another areas of mapecie somuth This moeibad promed quite I feepaund an satubody paint Fartin ta snya ting actul out But, ut . bigt alt the rabbit protna roto Thor good precipitatib kuin COM editation par e felt lasty odeat that their Then the state u brought together th the protein solated from my owo-B a precipitat formed, cosirmo ibe results Fnally in corroborate the amino acid com He obtamed 13 100tier und abused un cortic troa tb oyup gond from youn berdore Antartra mas pecific Just Deatoed (Tuu 2) ·540 Annals New York Academy of Sances thou. The third aurea in which grat drences have been made bs that of the fine structure of muscle. The Important fature of thb mort ure the at tempts to determine the locallation in the muscle fiber of proteins obtaloed in altro The fourth are in which interesting developments have occurred in the last fewr yan b in the #udy of chennkal change in muscle dong contraction lloc of this type has been done in various laboratoria, including those of 1 FH Blommaerts, Unrreruty of Culforna Sledal School Los Angeles, Calf Carlson s and my own in collaboration with A Casps of The Rockefeller Institute OY York NY There studies bayt rased interesting and per plenng questions, and nature oo far has refused to give direct answer to the question, what chemlal changes ocrur in angle twitcb? I pow return to the empler things which actually turn out to be not so umple that u to the proteins that an be studied in solution. The tem myouin has been in use for nearly one bundred yen, having been introduced in 1859 by Kubre although its original maning mua different from the senec in which it later ame to be wed Until the discoveries of Albert Szent Gyorgy and Straub in the arty torties, myo-un denoted the cabstance or ubstances that ould be crtra ted from ground muscle at a fairly bligh lonx strength about 05 JI but were mwoluble at low on trength L 1930 von Surult and Edor showed that auch an etinct posseto a very arong double refruction of flow indicating the presence of clongated, rather large molecules In the meantime the lactk acid theory of muscle contruction had been do arded and the phosphate-containing compounds that today are commonplace to biochemists come to the forefront of interest During the late thirties and early fortles several groups were able to show that myonn," tbe vlacous cttnct obtained from muscle rendue shon some remarkable factions with ATP It was dbscovered that on the one band, ATP has a profound effect on the phyucochemal properties of myouin The double reinction of for disappears, or considerably decreases, on addition of ATP and threads prepared by squrting myown into water contracted th ATP On the other hand, ATP itself lost phosphate group when it wu brought together with thb muscle atract These discoveries are associated with Needham dal Eogel bardt and Ljubunova, 'ood Szent-Gyurgy and hi co-workers' Thus it was apparent that ATP can affect the mechanical and physica chemkal properties of the muscle proteins and that the proteine, or the protein uit nu thought in those days an in tam ect enzymatkulty on ATP The tempting thought aroke that these mat purely be the reaction that underlike muscle contraction. The work of Lundeguard had already born that must coptnction is somebox linked to the break down d organic phosphate compound 1 x Bad reconsina ador VoIN It was won found that the wubstance re-nimel myosin-B by Srent yogi add hs colleaguet's not the only Interruing uulataire obtainable from the muscle redu it prebk Importet dercoscry bang on this proble u back by De los o cal lenguat the propunbon of the super ·Gergely Energy Utilization 513 T 3 PUT IT Fournom пи Сад і сту data + Myoen F-ketin (Strub-dommaerts) Foto from Ty 000-B Tropoo youn Iocohetoa o nablat antigo th utcunt kr 60 ron followed by adding ut TC for 10 boun The probation parlare contana 18 Laud 001 omby drony metbynamomethane buda A74 The antiqua su dlated 1 1 od than at ren coactatntion nnged from 003 mg/ml to 0 23 mg/ml two kinds of component, only one of which u the aumer of ATTEX ++ drew Szent-Gyorgyit has morggasted the term meromyodln for the product ob- ADtuned with trypen digestion, but in view of the fuct that now It la posible to obtain amilar but not quite enbaal fregident in many ther myn, such chymotrypur nod sabtılmın digortson, the term may have outlived its well From every point of view the fact that has A1Px ctivity meets to be the businer end of myouin, because it a not only an ATPue, but because it is abo able to react with uc tin this has been demonstrated mtb ultracentrs fuge by Alhaly od Soent-Gyorgy * Blolaular weighita I these fragmenta obtened by proteolytac digestion are still not settled definitely but I am in lined to accept the most recent maamonents btained by vanous methode in my laboratory' wuch are confirmed by Susan Lowey on th B logy Lab ontory of Harvard Univeaty Cambridge, dram and t ay that the mout likely values are about 320,000 fo the beavy and about 120,000 for the light fruction, so that one of ach of th two kind of fragments ppan on digestion of ono myouin molecule It seems that there must erat two farly distinct regions in the myoun molecule end us a reult of th ction ! proteolyt enzymes, these regions become separated and partly depraded parts the greater part of them 3 ando btedly left intact-appeur u what loosely bave been called meroyos There bas been come questooot whether the protcolytic eniyma that Jdille produce vurmus meromyoon mdeed act u proteolytic symbo brook " I think, u aulined to believe that they produce ther effect by more unknowo mechanion, but I should ny on tbe bune of our on nor and that of Gladner and roll that tho C terminal groups appearing after rypen und chymotrypon treatment are the very one that could be et>ec1ed if the crymer had acted a spectfic proteolytic enzymes Actin can be obtained from muscle rendu from which myou ther potable proton bave been atracted After this rude bu been trated with mild Jhah and ketone, the dry resklue (lat a tn ted with water the aqueous attract cootus the globular forma decim. As I mentioned curlier wits change the form on anolbar from that te haves in many respects us substances of high molecular acht are bote It apran behave, and bus been tamed F-uctin (F for fibros) Th intact Ada und alth ·544 Annals New Jork Academy of Saences b in some way Involved in this polytunnon reacbon, and it may be on intoyral part of the actin molecule The premce of ATI in etin you fint demonstrated by Straub and I cut and wouluently bry Lekim alan by Blommacris' It appears that globular tutia coulains approumstety one mole of ATI for cach mole of (ractin havoga moliculir mught of about 60,000 od polymerization ATT s transformed mto admoene diphosphate (ADP) - It seems that u libro ATP retential for mantuining the Gatm In . polymerlable form, but the F-acti much more stable. Kection of adonana ruk ATP od pyrophosphate. One of the most eriking properties of Futuns its ability to form a complex with myusin This soalled actomyosan has a mach higher rikouty than would correspond to the sum of the costics of the two components and has a marted double refraction of flow It ATT2X Lctivity in the presence of ligt far erred that a myouin. As m.catroned above in the arly phases of ibe more recent investigations on natacle proteins Kveral research groups found that mFort-B *buch we would Dow all natural actomyos, undergoos unhing physcochenkal changes in that its double reinction of flor doappars and its vecouly drops martedly when ATP added similar hangrs have been found to occur in light-cat toring studies * # During the last few years it has become an important problem to decide bat actually happens when ATT s added to other the natural or the reconstituted actomyosun The ngnlcance of the probleon les in the fact that it conceivable that the interaction between a tu myosin, or actomyoain, and ATP and the nou ons may givewa clue to the cents that take place in muscle of these proteins are indeed those responsible for mum ke contraction. The phenomena occurring in the actomyovn ATP geen depend very strongly on the monk Krength of the medrum At kom ons strength actomyounes bardly boluble and, under these addition, one an study either suspensions or thrads Addition of ATP ll produce a synterests of the kuspension or a contraction of the thread In fact, the obxton orginalh made by Albert Szent-Gyurgyt of ATP-induced a tomoun thread contraction 15 one of the most ruggestive piece of evidence favoring the view that actorvo- An ayolved in contraction in the whole muscle Hou ever at a higher ronk strength, about one-half molar for cumple ita powable to audy the previously mentioned changes in the souly light attening, double refriction of fion It , of courk, of great interest to knon what molecular chagos undertie the bull changes It has been named for 1 long time that ATP and pytop bophate and some other pubstance aux the disociation of actomyoun into actin and moon 2Јy оn орро в that, in fact, thus is the case my vrem bboxd on lgbt #cattering cyperiments that pbon deciaux in the wytrage molecular scight on addition of ATT and aboo on the receat xoth of Martonost in our laboratory By thing differential ammonium allate ulting-out techo que he was able thon that whereas an (tomtycun solutwe shomsone peal at about 30 per cent saturo Lon in the presence of pyrophaphate 180 xh peals are obtained od tber pooltloos lay losely correspond to the portion of the peaks or move and actin (TIGULL 1) uit blodeed the ae that ATP or pyrophosphate aino ·Gerech Energy Utilization 545 AD28010 AD 280 AXAS AXAS O NO PP •²MPLI 0.3 0.5 02 01 10 20 30 40 50 % AMMONRUM SULFATE SATURATION tool Dale walioutume de protonphate tallation chante (15)Chor 24 boere the pregnante contra ( od at 20,000 for 30 mu and the opel desty the grant un and in Bechoco DL pectropbotoncter 1180 m tbc duyOLLOO of the lomyan omplex and of the sculentually what beppers een at the bour slength prevailing in muscle the autumn studies may be of great wynfance in underst Oding some of the proposed moncle Dodels and would pronde a molecular mai buna of ourular contraction Tropposta Tropomjodi unother protestened to be in some ty uvolved m contraction Ius role a rather uncertun ultbough, on the bar olio amino acid ampoulou Betleya ongly greated us the name implien, that it mrbt be a buckling black of roon The da bas rently been elaborated upon with copoderable uma pamation and much bard or by Laki* He works in tropomy data and actin, together with some other not com pletely charm (enced protams, are the real budits of myouin Hori was I bel som add wenething about bar dacle procene ruodamentally the beart octomy oud THE xem to believe ey couch like the cletal one Vyoo from beat ucts, according to our own mayoreme molecular roght and shape u the skeletal ment, bu cially th th actm and ATP thom do Anting dutertoca protan, und the rektom Stern und hos ollespuest luve a different en bot the decrepancy hua pot yet been histu tonly resolved. There ars, honor to ports on hisch ring ATP bears myus dulas from thcktal counterpart mity Saood it paani be obxh more tbout xexe tbud thut of my oup It regard level of the proteoly1ic estant try pod bytootrypen ens, dius about to loves haghher to produce the punte viacouty stane In the ultracentnfogul sedintation dugom tho mejor componenu eppur but thus far atterapist Kepurate and isolate tbem baie mt bren aurreratul ·546 Annals New York Academy of Saences Rdersion of wodds Clycerdated muxl fibos ure as I rationed arkki halinay between the isolated 1* ti protan syRems and the living mode Weber und be collegues hare furnished, I think, very clear evidence for the close relation between ATP splitting and tenpon development in these fiber They have shown that various factors that affect the rate of ATP splitting almo affect the development of tension. This has been demongrated for a wide variety of factors such u temperature changes, ATT concentration, and willhydryl reagents it has also been shown that there is a close panllelam between the rate of putting and tenson development for a wide variety of nockoude tnpboxphates An important development Mus recognition of the fact that thert ure certain components in muscle that can effect relanton of the glycerinated muscle fiberu The obeervations of Jlersh" and Bendalle were the first ona to suggest that muscle itself contains a rela ang system. Subeequent work by Goodall and Szent-Gyorgy, and Lonand" appeared to pinpoint thus relaxing factor in the creatme pboxpbokinose system and Joos and Lorand" Leter succeeded in showing that the photoboedol pyruvate pyttyx kim, combina ton can aloo act as a relaxing factor The porable role ol myokimaxe has beco suggested by Bendall All of these system are phospbor transferring en rymes. For the first two the pbosphate donors are creatine phosphate and phoqbopyrivate, respectively for the last the phosphate donar i ATP The phosphate acceptor * ADP in all three aux These systems rexerable the original arsh factor so named by Bendall, to the requirement of digt sons for reaning activity and in being mhibited by Cat hom It was very tempting to speculate that these systems womehow act through the rephon phorylation of ADP bound to the muscle structure but all attempts to dem onstrate a mechanm of this type failed Another feature that has been demonstrated for most of these systems the fact that coocomltantly with the relating effect there occan en inhibition of the ATPuse of the fibers' Thu yppaan to be consistent with the iden daborated by lleber mentioned arlier that tendon development and ATPase activity go hand in hand The first æggestion that a simple phosphate transfer not all that might be involved came from Kumage and bbs colleagues, bo bowed that they needed at laut two proceo fractions to achieve a relaxing effect One of these contained myoknak the other appeared to be umiler fo its wdmentation chanctorutra to the particulate ATPok of Kelley and Jeyerhof • Tho observation has recently bero implified by Porlehl und Lorund den Hon ever Briggs und Partiehl have bat to me uppean to be good evideoke that the Deed for a repbispborylatmg syuem arises from the fact that most matigators of this problem bave ured rather thick fiber boodles. In this as the ATP CODcentration in the core of the fibet would be much lorur thun It has been shown by feet that in the peripheral parts and in the bath eod Holbach that the reluang effect depends on a taln nikal ATP concentration belox "hache there b no relaxation. Thus, a cordog to Briggs and Portreh!the repboepborylating Symons kae lo raste the ATP coo ·Gergely Energy Utiliation 547 centration in the central portion of the fiber to the required critical bod It appears that is single fiber och stoms a datina pagkolinax or mayo Linate huve no relaung effect in fact, they focrease the tission it ibe ATP Coordination is very box Brigpe bas now returned from Hicklebert to Harvard University and, meatly Kelder and I have collaborated with him in oor ditor to bring more light to this problem of the relawng factor Uang poste fibers, we ame to the cooleson that the rekaong sen connue of at least two components, neither of hubo any of ibe transpiramphorylating enzymes trentland cariker Hy eninſugar fruh murte etinut prepared with a stune-buffer solution At 35 000 inainn B obloured that by sets has no ormonly with relaxine buty I boxever the supernatant sudded the fibes relata If the rupernatant a dualyzed th dalyzate all aboo bare the ability to supplement the partulate fraction (IAT +) The complete muscle ettnct contains Lesre amount of the pranular portions for it a powable to take about one third of the moant present in the artgripally seth settinct and acthate it with an amount of the dualvate corresponding to the full volume of the original The deatzabt omponent appear to be a larly labile mubatance Another clue to to natare may be found in the fact that it is readdy abmarbed The fact that we were able to sperit tbe rearne synem tn these two fac tors ha ao opeoed up interesirag possibilities for aming further insigbt lato the relacrombip of ATPuke and relativo Althougt borter amoumus ar ith onlkt amounts the colartor r gramulo nhubut cayofibollar ATAU 10 ATPo tym (TABLE 5) qurement an be demonstrated also From ume to time pyropbosphet (PP)abo bas been implicated in the relo ang methanam, porek usarly by Bocket " lebe Wand Beartall in our Dinge het laused-op teto me nere able to show that If in concernatione cttrart ou chetoal TALL ETT OT TIETO COLOVI Roob na 04 ATP INDUCID Loker Trom 6 CUTAITO Snow Itt TI PODPD tou 50 SICOLE Crode act, I al Gno, uvut to i al code etik Supernatu 3500 ml Culo Apoulant Dalyate doplant Greta kod dalyzate Sorah Shut (@rtbrett sorra mbili mpat bed born atendi akad a to per cent phared stor CT date coad 2 VIE 3.VTP auto word wc ta 010, Wer 11 por contacto, o de una comes totekle Th 413 400 TX protected on the table us ondatruct het ces intaputo fotbed TJYL, und die Redundat de recorred to be unha Trouro oro wla poli petrol met soud to open more rtret ·548 Andals New York Academy of Saences TAL 3 Cmt TL Rumo S M ON VortNLAR ATTAC hat tapet Grurula, 01 ] Gatola, 4X dhl 01 m Grupula, ix al 01 ml + 7 I PP 2 IPP Gopala, 4x dl 01 ol + 01 ml duk med 35,000 spermatant 01 ml aperputant 77 0 81 0 60 The moodbrifer person to prepared by boeregalong in the long Blendor nabbut oktalende ths olema d 0063 boxcoate boller 75 follon ad by centraption t 1600 The Hoxtout darly showed to layers the bottom layer contained up broka cal and connective touc debrus, the per layer coolunmg the maink, orospended In the bed and the canopbon-responon procedunt repeated 3 more times The bu Abel per contained 16 No. The males and woperta tunt To popred a deronbed on the good a Awit the member spawa contuned 18 mg TWO ATP IN ucured out in the nechun ud for the propertion of the muola In total rolome al ml coctamg 3m Of ATP and 5 mji apso Tbe bexico contropad 01 ml of myolniler Dy to Olber addticos in ubated in the table The mermal phosphat pitting emoorted to 11 JI ID 5 min incubation at 25°C IDorpunkc pbapeut detarmmed det deproramabooth tncbloncet ud according to the Fl Le Sabbaron method N/in! of about 2 mJI does not by itself produe relation. If however PT added to the granules, it can replace the dalyable factor Clearly hom ever its not the active substance of the dulyate in view of the Lability of th uctul dlalyable component and its abeorption on charcoal Moreover IDorganic P Pare fohibits PP relawtion in the presence of granulos, but have the dolyable composent from muscle fully acthe PP bus umbar cffect on the ATTx of muscle fibrk in that concentrations of PP that by themselves bavo no effect on the ATPase uurity become inhibitory in the presence of much abounts of the particulate fnction that by themselves do not inhibit the ATP.x (TABLE 5) With this reloring system, us with the muscle proteins I db cumscd carlier the question of its physlologlal agaficance ulx ayı looma large in our minds As we gain more frought into the working of this relanng system ne shall what to Loow does it work in the intact mule bow as It incorporated into the general scheme of csatation and relation of the developerent and the deany of the active sate? Ile should like to know how it loculued in the muscle that is the some question se ush in connection -ith thone muscle protes about which we bave much more detaled Lowledge at this time flon ever while we annot mer these questions with respect to the relaang fuctor I think we have much more information about the structural proteina docuddarller ondar srce on trochu As regards the problem of the localiza tion of the structural proteins in the musclo fiber the model developed by Hon und Hurley sons to be apported by the evidence ultbough karne ·Gergely Energy Uullination 549 Investigators, induding Hodge * think that other modete are more plausible. Podolky che here in tho monograph has shown that i ther the cbakal" mode imoving shorteomg of protein filaments, or the interdiciutiny one based on the relatne motion f protein chuim whore onfiguration doa not change, coulde ount for the ntc of art Liberation obserred by HIJL The proponents of both types of modeto have akcioned that myoss and actin uwe Lnon the protein in volutos, eo etia in the recte filter er and Janbul receatly reported some experienta with thoorcocent amit Holt bodes that tont glance might render the supposition unterubk. Thoxe inyoligaton prepared noorement actibodies galost the po-called mamy okres and found that the inubody guinst the heavy cotopobat reacto only with the middle of the band, the maled I conc, hertu the other eppern to be bound to the rest of the A-bend. I do not believe that it can be meld at the stage that this s evidence for the balance in fatact macle of theme ants smaller than myran If the components reacting with the oth bodies were ondeed, closely related to the fragments springt which the anth bodzes nett obtained and f, indeed, the smaller unut ensted La Dusche in weddy wanted regions, it would be necessary to decand out whole concept of the myoon molecule This all rather nem atmal and I do not think that we have sufficient credence for truly chical docion, but I want that t is quite oncevable that the structural relationshupa enting to mbacke may interfere with the nation of the antibodies and protdra such a way that the resulting patteto s the one dexnbed by Blenbal and Holter It may be more than coincidence that the anti-H-meromyouin rezct only in that part of the arcome where, curding to the Horey Hansen picture there 3 do actu Remembering that H-meromyoan reacts * fibre sith artra, at may be powable that in th rest of the A-band the rende boa preventa th combination with the antibody I would urge great caution in the use of terminology Bleromyouin labo arad a generic term for renous ingment produced from mypun by proteolyik there is musiche because of the chemka chengas brought about by the proteo tyme Strictly speaking the Inteotannot be delical with any lytic anime Thus, if units muller than myoon etatu dlm intact macle they cannot be dentified with any krd of meromyoain consequently the bending to purcle of unubodan punt the proteolytic meronyoulon cannot directly be uued us mes for locating Kructural components The Dot mere quabbling I feel strongly that to dostand the polts a qute pombly to introduct artisanal complication to the already complicated picture If we do accept t lax for the parpone of a fument, the model put forward * contracting met both by Hurley rod Hammon,' re must regard touche netin of our inom ledge of the rol & compoDedi 12 lb ind in terms of Threrla of the powobutky afforded ly the Kructure of the muscle aber muone ke tan lo the post het my ongidered the contraction as based on some hange in th automjon mokrak or I some othen he n I been quatc u plicite that they stalnly ha t condered changes in the shape and configuration of swe materskes us responsible for contractoa.

·550 Annals Ney York Academy of Sciences For some time, however one difficulty has been pointed out that the fact that in X ny studia no dgnificant changes could be detected among related contacted ind stretched muscle If a grou couling or upcouling occun in the contraction cycle it should be viable in the diinction dlagrums. How ever the modd proposed by Huxley and Hannoo produces an entirely different atuation. Het re have a longitudinal displacement of molecules with respect to ach other without permanent danges in the molecular configur tions of the molecules themselves In recent revich A Harley bu ar plored the implications of the Huxley Hanson model and hos utempted to show that soch , model rould lad to consequenca consetent with the known properties of muscle us a bole toewhere in this monograph Podolsly discursen pome moro general features of thermodynamic thoones of muscle confraction I now propose to discus the implications of the Hurley Hanson model in terms of known chemical and ensymatic properties of the muscle proteins It appears to me that i contraction phare based on the combination of myosun and actin involving sulfhydryl groups of myoma rould be consistent with the knonn tendency of myogn and acun to combinc. If u I arlier Arrested one could take the reaction of actin und myoun in volution with each other and with ATP us an Indacation of what happens in muscle, one would nec that, H ATP wus romehow aloned to combine with the relevant ate, it would cause d.ocnation and, hence, in terms of this model relaxation As long u ATP could be present around the atas involved in the combination with actiq, the actin and myown would remain dissociated and no contaction would occur There s good correlation between the effect of sulfhydryl reagents on the tengon exerted by glyceridated fibers and the requiremenu of the picture in which the blocking of axta altos would be opected to land to docuation and relation The relaang system, by mhibiting ATP. ac tivity tends to increase the number of ATP molecules per the ata javolved both in splitting of ATT and in the combination with actm The perintenet of ATP in the related state would also be favored by the fact that in the prer ence of magnesium, myosun desocuted from actin has very low Allu activity Era by sowa o mundo contractor I now turn to some problems of the energetics of muscle. The first question that comes to mind a tho Is ATP Involved in the elementary crent of muscle contraction? Amers to thos question bave generated considerable Interest, some but, aod pomably som light, thanks to the work of blompact, Fleckengtan, Krebs, and others wen Tbeir wort aiggests, fins faac that during a angle twitch or even during • short tetans, no detectable changes occur in ATP ADP inorganx phosphate, or creatine phosphate. Ia my oro mind there is no doubt that Blom maerts areful experiments bave shown that, us far as ATP and ADP art conxemned one an ophantatingly ay that upder cooditxons where repbophorylation both acrobkc and anaerobic, not inhibited Do changes occur Oning to technical difficulties I am not certain that one can say but the data really exclude changes in the creatine phosphate kvelunce the total cypacted change would be about one quarter to one half \l/gram of musle ·Gergely Energy Utilization 551 and the total creatine coalcat u 10 2 The data of Technen und ba colinarik, due to the use of different fachnique, male the lack of change in the creatise phosphate level more convincing Honever ma neither of tbexe aeruments as the glycolytic cycle inhibited, and I think it would be entreisely hamardow to uy that phosphate inander could not laure curred Another difficulty with a capetimes the type is the method of hauon Agun, I think Voromateru bas probably gone u far u poble na mabing ATC that boucles do Boi doirat at the moment they are fuced by freezing Ilon ever it is one thing to say that there is no valor contraction and quic arot bet to nume that no hetkel a tivation keading to changes in the pbophut daininition tus ocurred Spectroscopre maastiremenu ofbe Type tasted out in Chan and has co-ority" may eventually tbron more light on this questiod although at the moment their experiments are not concave There who goeation that in the banget kesin of mitchen ATT doar dup le could are the clown Kork of Lundeguard and, more recently the Morb the rockerscen e do. Recently both Capol and mywell and alone Carison et al have been that the fir suberance to show change a creature phosphate, and there on lusty good lucernity beiween the duntion of simpla tron and the change in the chatne poomphate level Therefore by mag the mashenuatkal argoment of continuity it ould seems that we could define an unoant of creatine plohet rrespondmg to * nogle twitch and that one and are also that the wote etwount must disappear m the first twitch the knen, HDCO DO break in the curve ud tertanke It seems to me that whether ATP dappearance ao be shown mat mode im ich o hardly one of the most fundamental queRICE me that there an un nonne or Ungpetted suberance « XP that plays an important part in the straction It may mt either directly directly ATP may be able to repliosphorylate thua substance very rapidly so that even in the tire thu h of a Kia change would be dete table Alternatrvehy it may not be able to repompborviale speedly in nbach (Rm the firut twitch a to perbapa, sul not be accompanied by changaa n ay known substance Honder the stuuton could burn d 477 reacted womedutely and run I was repboxphorylated very rapidly Therefore I think that the runl queerns are of mechanm and Dos Hombrotretry word that belie anno nude v KTY Only as Lundmarte exploring the mesown territory o the cheska mechi ung twitch to the purpose of th fosion docton I shall werde mod I (bunk moet people was egres, that the ultimal source of the entry of our le contraction COOKI of some besnikTKY prophetas, the ball of which are present at ATP and content phosphate to be most general terms the problem of gergy otillabon node of thermodynamice, in the count that the firut berat thermodynamos » not permit ist producer nach einen rotapand to the total chemical carey prruett tbe mood 4* u napomc further inte tuont la tento debdebey us that only the free energy di chemical reaction Sinct ratually the liberation of weby If directhy tun kun lable for mechanikat work ·552 Annals New York Academy of Sciences from the high-energy phosphate compounds Lovolves the hyrolysis of one or more phosphate boods, the obvious thing to do u to look for i system that an put phosphate bonds Vost people worlure in the field of murcle ren search bave concluded sot unnaturally that the ATPase activity of myunin and actoryou cannot be considered uudental but that Ibey must have an important role in the liberation of energy for ontruction Certun dif ficulties here beca nbed in tbus connection by Slotmatta, ho baus pointed out that the ATTıke activity of myoun us too low to account for the phosphate liberation in livlog murcle. Hon over the recent work of Morala and hu collaguas" han sbomo that sobatances that apparently have no knoma role In the metabolism d muscle, fuchu aminoethylbothiouronium can conoderably increux the ATAX activ ty of muscle. Therefore, by a spurning the costence of similar unknown action in muscle or by asuming that the Itnictural relations hare ugnificant effects on the ATPase activity the pp could be considerably Dioned In connection with the latter point it should be noted that the activity of punified cytochrome C s much lower in a reconstructed system than in the intact tysten The 'how of the convernor of the phoqphate bond energy is of the greates theoretial interest in the problema of bat might be called chemicomechanical coupling The theories could be dnded into two groups clanacal ones and perbape, those that might be called quantum-mechanical theoriesIn the latter group Albert Szent-Gyorgyi s one of the propeen Of course, it should be erophasized that even the clanical hemuts of today are cognisabi of the trse of quantum mechanics the most clar cal-looking organic chemical re action can be described in term of delocalized rital within vinou orgina molecules The effect of certain substituents on the activity of aromatic copounds has been most clegantly disclosed, for euample by Longuet liggins For the purpose of this discharon bometer he can malc e f concept rxh 1.1 group transfer charge effects, change in phare due to entropu ta on, and other Stent-Gyorg? advocates the consideration of the pomubuity of energy transfer throughout the brakogueal fabric wmilar 10 that acuting in an electnc conductor or semiconductor with an elettron gas If the structural View Alggested by the experiments of Holtzer and Varshall Bere to prevail, some poch mechabam would be almost evitable Race the ATT glitting would occur ta patial sparation from the rest of the contractile mechanson Even in this ar one would eventually find it decorary to make use of the classical concepts in descabing tbe actual mechanism of contraction Justi, m describing the working of electrc motor for comple in which the transfer d energy in the more proceeds according to the quantum meclunk a concepts of the electron gas, & must be made of dancic mechankal concept and devkas, much u momentum, fears, and alles It o pomible hon at that in muscle we may eventually urlie at i description that can depende with all localized concepts mich us aborteoing of bonds, folding of molecuks, and shufting of chain iod, u Surdt-Gyorgt grests, there may be abole sake change in the water inkture so that the hole tructure could collapse if the morty womebow cither laid out or were put into it ·In the coming 1050 8 Nstert 1971 129 15 Tom Pror Most Nir B Gergely Enero Utilization I think that here ne have seached the rolu set by the principle of comple 553 Deplanty and that only (ude all indicate cu lutore courk. son the vzhues of some of our obterrubks, wing the language of quntum mechanics will have changed froen umslamp to "aharp kad berean today kemat pul in cms of a probability dostobution of theories, nc may then antre at a ertainty responding to an ergenster Romer 1 Bom, 1919 best I ostae Mbomber Bakalat 2 Hul, ESTAS 210 1946 Bt Wed the u: 174 1 sn GOYTA Borneo dedetox Isen ATTI GLA 1949 Door Han 100 Tew York, NY WR, H 1933 Ilang Lectro Ser 41 37 6 KILI 18.50 Arcb Anal Payo 749 7 SI O tom, 194 Acta no Strod Berpol 23 7b Sr Gideon, A 1946 Moncler Contraction Aarden Tree Yey lock on Mou HAFDAL 1930 j Had Chaos 87,315 9 LINDAAARD E 103 Baadero Z H7 31 10 \rnX) STY DVE YOMAX ASC Law 766 1941 11 [VOULARDT VIIM N Louw Vatant 1141 668 12 Bu 31 IBU &F Guy 1957 Natare 17). 818 13 SEXT G2 1931 J Biol Cher 19: 367 14 Won, 1056 Bodhin e Bropby, kte it 3-45 Ilmu PHBS 1 Count ! Sau & FLORAL (bem Sox tbar UIT 1937 10 KAN DR i Roca P S MADE & Lus, 1954 106 Horn HARIAL 1956 Pror People Mac Boston, How 16 GroeJRA Martinus 199 Falentom Proc 27 228 35 17 LAUK in CALDO 1945 Matwo 173. * I HOLTON A Slon 1956 y lo Chan Soc 1953 J&H komun 1957 Proe Cool Mwache Contraction Toky Jepun TI FHI&B B ALDAD 1957 Sacace 13 1201 21 Grote 1990 fodeninon Proe 176 21 21 IGOR VI ID KARISTA 1955 J Bal Charles su 103 Gnom 2 GROEL Bo Chre DOC 24 St Go 1090 Arct Broco e Beophy 305 25 VoortOK HR 1953 Buopiya Sor 26 VUSAL FLORA Biol Chuo 211 27 Crrant 1 Kola, 15 RAI VA 10:58 Proc Baby Sur 40 20 GLAD Four 1019 Ba C l 2t 1050 Buches et plan bu 455 Luk Bonn lun 1050 Joer Phyno dun 31 VODITI, VI 1951 Ba Cha in $.99 32 ostvari, FH 1952 Bu be Ik. 45 BLUX IV Vous Job And Barbera Beopets us no 3 Orbe 1958 Book Chao 911 35 BLXXXF Joku 1913 in Bocko Bad) U 2* * Bar 1919 Hochh771 37 La IH Pror Cool was sixto Tokyo, Jyour 3 GT IN LONLA 1957 Federation Pror 16. 103 1946 lader on toe 1 90 91724 TIL 40 Buxbo LIS 41 R 19 12 11 HBB TO! Na 1 1005 to Natare ITL & +) (patium HIL 1953 Vature in 1181 thuctio et Baby Lu #4 #51 + Muo, & lot v to DENDALS, JR 1931 Proc Roy Soc Loodo *09 Arch Bochem Beopby 19 G Nora 1! Goto m RIO 105 1857 ·554 Annals New York Academy of Saences +7 BADALL JR 1953 Phymol 11 232 49 KUKA,,S D&F Toma Loss Start 171106 19 KILIY TY TV ZOJ llam 1950 10 391 50 ParaLH 1957 Bochun a Biophy Act # 373 31 Lown, L J Homu LC Book 1951 Proc Coof Dorcar Contraction Toby Jepun 57 B poo, FN&H TOTEL 1957 Bochum et Biophy Act 164 536 Wom, H H&R HOCA 1993 Bloctum at Brophy Acta 11: 160 Jb GPOOLY JI N Bimos & G KALDOI Abet ttb Lutero Coop Biochem & S4 Born, E 1951 Am J Phymal 151 776 35 WOD, HH 1951 Z Eld trochem W S11 56 BETALL JR 1953 N to 172. 586 57 Harton, JHorn 1955 Sy opodm Soe Lupul Dal 223 U HIQDOL, A 1956_ Brophyz Boden Cytol 4(3): 131 $9 HUXLEY AF 1957 Propor Brophy 1 257 60 VOTI, SV FHN 1954 Netora 176 1081 61 Monum, FIN 1953 Am J Phy wol 101s 67 TLICHO, A JANKL, REDE & H A Kur 1954 Nature 17. 1081 3 CUC, B ' cil Com 1957 Nam H 11 61 JOM, r&B C 1957 Federation Proc 16 68 65 EALLOY, D F San&T C JNo 1958 Proc Biophy Con 46 66 DINAMIT, FH1 1950 Simecular Contration IncroXDT low lank NY 67 Mon, IFAJ OO, H L HATTER CRANIU 1957 And Bo chem Biophy 11 68 SLATI, E C. 1919 Brochem J H 305 69 Sott G ON A 1941 SOCDC 609 690 Lorot Hot, H1 C 1950 J Chen Phys 13. 283 10 SIET-G Oray, A O 1935 Adiunc id Ensymbol 16 313 71 Pour SV 1956 Phyol Rey #: 1 77 JOLALEL, MIF J Bom, J J Blow & TL HL 1955 Plywol Res 350 475 73 ton, HH Am Rer Biochem $ 667 74 MOLLS & STEIX 1951 J Biol ChemIA 663 75 CAY DFAM DILLO &RE DA 1958 Ntart 162 721 1954 ·The folloret calculations are rubd only as long as the energy prodocod, tude from best only the mechanical rock espended by the heart in overcoming resistance I tbc prctance of constant attaml work, the ciiciency of the busting ta Catbetont of tbc coronary unus hus made ft pour to twy w have codicated that fatty acids furnush the grate conti loution to the ti the contnbutro of arbohydrata od non-carbohydrato to total ECT In the lasting date, the relationship between The haut an age all Rubanto circuit y individual gibutant utilared deed Stonpe of foodstuff by the beurt om I cumpl arbobydnta ure Kored us dycogen und fatty adds a neutralin catan abnormal conditions, bone Blycolyth of the bart mat play a role may prodoction According to expectation, this occuc the orygon rupply to th bart muscle diramare, u for ek kr ardu infarction and following ute of the curomary drculation these oodatxona, lactate and pyruvale conceptntlons in coronary stawu Thank reported the pepo uported by ſtod in the club H270) Polk Halk Star, Berbeda dpd the W}x TRDIAL ELEICIENCY RJ Bung and G Vxbal Diperet af AT A uw laron Scheme Vana and the Halbtage I or Wax fort i rum du lese St L, Y Futbfol adherence to the lopk of heart officiency would lead to a dirty of dynamk sents, fxh us rostana flon and pro paragraphe, hon entr ure restrated t SETH of txo factors that aucunt the calculauon of myocandal checkbcy anaerobic bretabolar (plscola and otygen consumption of the onbenung hant. The final portions are paper are denotedt die son of myocardial party utilization. The efficiency of the heart can be determined by the nth of the there's equrulent of the work performed the heat liberated. Therefore, la be indirectly cakubated by ompaning ibe work performed with the my court orygen age To former as sytuonymous * th energy atllation the later Prents total entry production Determinations of myoardial efficace from th Orygen consumption of the heart annot be mado unlem one line that anaerobkc garalyses in bart muscles abonent An additional dische 10 the cakulawo of myocardial office ency arties from the fact that is the shendre end pulmonary circulation and to traparting moment tom to blood from Therefore i Loom ledge of the grypen consumption of metabolism of the bart in closed chest animal and in man DOS production of th bant production about 1 to 3 blood, and in groeral the imount of on ju tetal concentntion. There is no evidence of glycolyma in the normally beating heart that uced those in urtetal blood Anterobre glycoly tt Gant V Vedral Resourb food, the Ann Flart Amateur und ih. Toberto Industry Roanh Cortes, No SNY There wa tie retrebko, pront ·556 Annals New York Academy of Sciences TANI 1 OL la nunt) Terkeca (nut Arrested dosbarth Bating dog betrt Batrog med baut IO O 34 0 254 It 3 the penod of shemu and ADOTETILA B Dot too ettene (low than 2 to 3 horn) The second factor that plays an important role in determining myocardial efficacy is the orygen consumption of the artaged heart It hu licen demonstrated that the otygen umge of the beart arated with potanum chlorde varla betwetu 20 to 35 per cent of that of the naturally perfused hart * * This finding is in agreement with results obtained by Greg! and Sabaton A compartoon may be made between the consumption of the disco and that of the whole utested heart 12 SIX Thus necesitata CONVEtwan of myocardal O, bage into units of dry waght, which can be accomplished by a factor 5 22 reprehenting the relationship between dry and wet weight of time mica The results of these akulations are presented in TABLE 1 Studios from the laboratory bave shond that there is no agmficant difference between the otygen conginpuno od the beart beating with empty valcles, the fibrillating heart, and the beert atented with potupum chloride In contras Lorbers has found the oxygen consumption of the fibrillating beurt to be greater than that of the arrested or of the normally beating heart powable that the high myocardul otyger unge reported by Lorber retults from the fact that in bo preparaton the chambers of the heart are filled with blood thadore, the tension and length of the muscle fibers are greater than in the experiments reported from thu laboratory where the cardiac cbambini were empty. The relatively high orygen coupuon of the urrehed per fused beart Decomutato a re-evaluation of previously publshed data on myo cardlal efficiency Such alculations can Dow be made from the difference in myocardial O, unge between the naturally beating and the arreked dog beart (mean O, wge of the beating heart is 25 11 ml/min mean, of the arrested bart $ 4.83 mlinin.) Consequently the O, consumption of the latter 19 4 per cent of that of the beating beart Actordingly 19 4 per cent of the total O, confophion of the left ventricle is mod for bagal metabolic proctext exclusive of soch. The efficiency of muscular activity of the bart can there fort be obtained u follor, Work of bart o coobumption/mio X 2039 X 0806 where 2 059 a the mergy squalent (lg ml) of 1 ml Oat in RQ of 082 and 0 806 I the fraction of wed in the contractile work of the hart only According to these akulations, myocardial efficiency s approximately 37 1T coat in the dog ud 39 per cent in mo

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