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Otto Fritz Meyerhof and the Elucidation of the Glycolytic Pathway

Open AccessPublished:January 28, 2005DOI:https://doi.org/10.1016/S0021-9258(20)76366-0
      The Equilibria of Isomerase and Aldolase, and the Problem of thePhosphorylation of Glyceraldehyde Phosphate (Meyerhof, O., andJunowicz-Kocholaty, R. (1943) J. Biol. Chem. 149,71-92)
      Figure thumbnail gr1
      Otto F. Meyerhof. Photo courtesy of the National Library of Medicine.
      The Origin of the Reaction of Harden and Young in Cell-free AlcoholicFermentation (Meyerhof, O. (1945) J. Biol. Chem. 157,105-120)
      The Mechanism of the Oxidative Reaction in Fermentation (Meyerhof, O.and Oesper, P. (1947) J. Biol. Chem. 170,1-22)
      The elucidation of the glycolytic pathway, the process whereby glucose isconverted into pyruvate and ATP, began in 1860 when Louis Pasteur observedthat microorganisms were responsible for fermentation. Several years later, in1897, Eduard Buchner made the significant discovery that cell-free extractscould carry out fermentation. The next important contribution was from ArthurHarden and William Young in 1905. They realized that inorganic phosphate wasnecessary for glycolysis and that fermentation requires the presence of both aheat-labile component they called “zymase” and a low molecularweight, heat-stable fraction called “cozymase.” (It was latershown that zymase contains a number of enzymes whereas cozymase consists ofmetal ions, ATP, ADP, and coenzymes such as NAD.) Building on these initialobservations, the complete glycolytic pathway was elucidated by 1940 by thecombined efforts of several scientists including Otto Fritz Meyerhof(1884-1951).
      Meyerhof was born in Hanover, Germany and grew up in Berlin. In 1909, hegraduated as a doctor of medicine from the University of Heidelberg. Aroundthis time, Ludolf von Krehl was building a small research program onmetabolism at the University of Heidelberg Medical Clinic, and he offeredMeyerhof a position in his laboratory. There, Meyerhof met Otto Warburg whoseinnovative ideas and confident approach inspired him to focus his career onphysiologicalchemistry.
      All biographical information on Otto Fritz Meyerhof was taken from Ref.

      States, D. M. Otto Meyerhof and the Physiology Department: the Birth of Modern Biochemistry. A History of the Max Planck Institute for Medical Research (http://sun0.mpimf-heidelberg.mpg.de/History/Meyerhof.html)

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      1All biographical information on Otto Fritz Meyerhof was taken from Ref.

      States, D. M. Otto Meyerhof and the Physiology Department: the Birth of Modern Biochemistry. A History of the Max Planck Institute for Medical Research (http://sun0.mpimf-heidelberg.mpg.de/History/Meyerhof.html)

      .
      In 1912, Meyerhof took a position at the University of Kiel. A year later,he delivered a lecture on the energetics of living cells, one of the veryfirst adaptations of the physical laws of thermodynamics to physiologicalchemistry. Meyerhof had recognized that after energy is input as food it istransformed through a series of intermediate steps and finally dissipated asheat. He soon began using muscle to look at energy transformations andchemical changes during cellular function. Meyerhof was also interested inanalogies between oxygen respiration in muscle and alcoholic fermentation inyeast and proved, in 1918, that the coenzymes involved in lactic acidproduction were the same as the yeast coenzymes discovered by Harden andYoung, revealing an underlying unity in biochemistry.
      Soon after World War I, Meyerhof began collaborating with Archibald VivianHill who was investigating heat production in muscle. The pair worked todecipher metabolism in terms of heat development, mechanical work, andcellular chemical reactions. Meyerhof determined that glycogen is converted tolactic acid in the absence of oxygen and showed that in the presence of oxygenonly a small portion of lactic acid is oxidized and the rest is converted backto glycogen. This discovery of the lactic acid cycle provided the firstevidence of the cyclical nature of energy transformation in cells. Theseresults also confirmed and extended Louis Pasteur's theory (now called thePasteur-Meyerhof effect) that less glycogen is consumed in muscle metabolismin the presence of oxygen than in its absence. Meyerhof and Hill won the NobelPrize in Physiology or Medicine in 1922 for their analysis of the lactic acidcycle and its relation to respiration.
      Two years after wining the Nobel Prize, Meyerhof joined the Kaiser WilhelmInstitutes in Berlin-Dahlem. Then, in 1929, he took charge of the newlyfounded Kaiser Wilhelm Institute for Medical Research at Heidelberg.
      By this time, it was clear that glycolysis was far more complicated thananyone had imagined. The sheer number of components and their short livednature made the task of sorting out the pathway daunting. However, during histime at Heidelberg, Meyerhof's group was extremely successful at breaking downglycolysis into its many separate components. In 1932, Meyerhof made the firstassociations between the uptake of phosphate during the breakdown ofcarbohydrates to lactic acid and the splitting of ATP. By 1934, Kurt Lohmannin Meyerhof's laboratory provided direct evidence that ATP synthesis was thebyproduct of utilization of glucose. Lohmann also established that creatinephosphate is an energy source for ATP phosphorylation, which led Meyerh of tothe conclusion that the energy release from ATP hydrolysis was the primaryevent leading to muscle contraction.
      By the 1930s Meyerhof had managed to isolate and purify the co-enzymesinvolved in the conversion of glycogen to lactic acid and had reconstructedthe main steps of this set of reactions in cell-free solution. All in all,Meyerhof's group discovered more than one-third of the enzymes involved inglycolysis. In 1932, Gustav Embden constructed a detailed proposal forreaction sequences for almost the entire glycolytic pathway. Over the next 5years, Meyerhof, along with Warburg, Jacob Parnas, Carl Neuberg, Gerti andKarl Cori, and Hans von Euler worked out the details of glycolysis, which isoften referred to as the Embden-Meyerhof pathway.
      With Adolf Hitler's rise to power, Meyerhof left Germany in 1938 and becamedirector of the Institut de Biologie Physiochimique in Paris. In 1940, whenthe Nazis invaded France, Meyerhof fled to the United States where the post ofResearch Professor of Physiological Chemistry was created for him by theUniversity of Pennsylvania and the Rockefeller Foundation. He remained atPennsylvania where he continued to study metabolism until his death. The threeJournal of Biological Chemistry (JBC) Classics reprinted here arefrom Meyerhof's time at Pennsylvania.
      The first paper deals with one of the intermediate reactions that occurs inglycolysis: the splitting of hexose diphosphate (now known as fructose1,6-bisphosphate) into two triose phosphate isomers, glyceraldehyde3-phosphate and dihydroxyacetone phosphate, by zymo-hexase(fructose-1,6-bisphosphate aldolase). Triose-phosphate isomerase then convertsdihydroxyacetone phosphate into glyceraldehyde 3-phosphate. In the next stepof glycolysis, glyceraldehyde 3-phosphate is oxidized and phosphorylated tobecome 1,3-diphosphoglyceric acid. Warburg and Christian(
      • Warburg O.
      • Christian W.
      ,
      • Warburg O.
      • Christian W.
      ) and Negelein and Brömel(
      • Negelein E.
      • Brömel H.
      ,
      • Negelein E.
      • Brömel H.
      ) proposed that this stepoccurs through the intermediate 1,3-diphosphoglyceraldehyde with the aid of anoxidizing enzyme and cozymase. If this were true, then inorganic phosphatecould be used to remove glyceraldehyde 3-phosphate from the hexose diphosphatereaction.
      To investigate this matter further, Meyerhof and Renate Junowicz-Kocholatyredetermined the equilibrium constant for the isomerase and aldolase reactionsin the presence and absence of inorganic phosphate, cozymase, and Warburg'soxidizing enzyme. They found that their values agreed with those previouslydetermined and that equilibrium is not influenced by the presence of inorganicphosphate, cozymase, or Warburg's enzyme. They were also unable to detect theformation of any substance that would break down into glyceraldehyde phosphateand phosphate, prompting them to write that Warburg's claims of adiphosphoglyceraldehyde intermediate may have been“premature.”
      The second Classic deals with the next two steps of glycolysis shown asReactions 1 and2.
      Glyceraldehyde3phosphate+phosphate+cozymase1,3diphosphoglyceric acid+dihydrocozymase
      Reaction 1


      1,3Diphosphoglyceric acid+ADP3phosphoglyceric acid+ATP
      Reaction 2


      Harden and Young stated that during fermentation, one sugar molecule isfermented to CO2 and alcohol while a second is esterfied to hexosediphosphate (
      • Harden A.
      • Young W.J.
      ). In a cell-freesystem, this reaction can be divided into two phases, a rapid “phosphateperiod” and a slower phase that depends on the rate of hexosediphosphate fermentation. Meyerhof proposed that the rate of the hexosediphosphate reaction was much slower in cell-free systems than in live yeastbecause the majority of the enzyme needed to split ATP, adenylpyrophosphatase(apyrase), was lost during the extraction process. He backed up his claim bystudying the distribution of apyrase in the yeast cell and showing that itremains mainly with solid elements that are not used in cell-free systems.Meyerhof also purified apyrase from potatoes and added it to cell-freepreparations to prove that it raises the rate of hexose diphosphatefermentation.
      The final JBC Classic revisits the phosphorylation of glyceraldehyde3-phosphate and its subsequent oxidation. In this paper, Meyerhof and PeterOesper use a Beckman spectrophotometer to follow the reaction and providefurther proof that a diphosphoglyceric aldehyde intermediate does not exist.They also alter the equation for this step of glycolysis to reflect the factthat the reduction of cozymase is accompanied by the formation of anH+ ion.

      References

        • Warburg O.
        • Christian W.
        Biochem. Z. 1939; 301: 201
        • Warburg O.
        • Christian W.
        Biochem. Z. 1939; 303: 40
        • Negelein E.
        • Brömel H.
        Biochem. Z. 1939; 301: 135
        • Negelein E.
        • Brömel H.
        Biochem. Z. 1939; 303: 132
        • Harden A.
        • Young W.J.
        Proc. R. Soc. Lond. Ser. B Biol. Sci. 1908; 80: 299
      1. States, D. M. Otto Meyerhof and the Physiology Department: the Birth of Modern Biochemistry. A History of the Max Planck Institute for Medical Research (http://sun0.mpimf-heidelberg.mpg.de/History/Meyerhof.html)