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. 1965 Feb;94(2):353–367. doi: 10.1042/bj0940353

The oxidation of d- and l-glycerate by rat liver

P D Dawkins 1, F Dickens 1
PMCID: PMC1206517  PMID: 14346088

Abstract

1. The interconversion of hydroxypyruvate and l-glycerate in the presence of NAD and rat-liver l-lactate dehydrogenase has been demonstrated. Michaelis constants for these substrates together with an equilibrium constant have been determined and compared with those for pyruvate and l-lactate. 2. The presence of d-glycerate dehydrogenase in rat liver has been confirmed and the enzyme has been purified 16–20-fold from the supernatant fraction of a homogenate, when it is free of l-lactate dehydrogenase, with a 23–29% recovery. The enzyme catalyses the interconversion of hydroxypyruvate and d-glycerate in the presence of either NAD or NADP with almost equal efficiency. d-Glycerate dehydrogenase also catalyses the reduction of glyoxylate, but is distinct from l-lactate dehydrogenase in that it fails to act on pyruvate, d-lactate or l-lactate. The enzyme is strongly dependent on free thiol groups, as shown by inhibition with p-chloromercuribenzoate, and in the presence of sodium chloride the reduction of hydroxypyruvate is activated. Michaelis constants for these substrates of d-glycerate dehydrogenase and an equilibrium constant for the NAD-catalysed reaction have been calculated. 3. An explanation for the lowered Vmax. with d-glycerate as compared with dl-glycerate for the rabbit-kidney d-α-hydroxy acid dehydrogenase has been proposed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BARTLETT G. R. Colorimetric assay methods for free and phosphorylated glyceric acids. J Biol Chem. 1959 Mar;234(3):469–471. [PubMed] [Google Scholar]
  2. CAVALLINI D., FRONTALI N., TOSCHI G. Keto-acid content of human blood and urine. Nature. 1949 Nov 5;164(4175):792–792. doi: 10.1038/164792b0. [DOI] [PubMed] [Google Scholar]
  3. COWGILL R. W. Paper chromatographic separation of 2-phosphoglyceric acid and 3-phosphoglyceric acid. Biochim Biophys Acta. 1955 Apr;16(4):614–614. doi: 10.1016/0006-3002(55)90300-0. [DOI] [PubMed] [Google Scholar]
  4. DE LA HABA G., LEDER I. G., RACKER E. Crystalline transketolase from bakers' yeast: isolation and properties. J Biol Chem. 1955 May;214(1):409–426. [PubMed] [Google Scholar]
  5. DEN H., ROBINSON W. G., COON M. J. Enzymatic conversion of beta-hydroxypropionate to malonic semialdehyde. J Biol Chem. 1959 Jul;234(7):1666–1671. [PubMed] [Google Scholar]
  6. DICKENS F., WILLIAMSON D. H. Metabolism of D- and L-[3-14C]glycerate by liver tissue of the rat. Biochem J. 1960 Nov;77:356–363. doi: 10.1042/bj0770356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DICKENS F., WILLIAMSON D. H. Studies on the metabolism of hydroxypyruvate in animal tissues. Biochem J. 1959 Jul;72:496–508. doi: 10.1042/bj0720496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DICKENS F., WILLIAMSON D. H. The determination of hydroxypyruvate and glycolaldehyde. Biochem J. 1958 Jan;68(1):84–88. doi: 10.1042/bj0680084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DICKENS F., WILLIAMSON D. H. The preparation and properties of lithium hydroxypyruvate and hydroxypyruvic acid. Biochem J. 1958 Jan;68(1):74–81. doi: 10.1042/bj0680074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. EL HAWARY M. F. S., THOMPSON R. H. S. Separation and estimation of blood keto acids by paper chromatography. Biochem J. 1953 Feb;53(3):340–347. doi: 10.1042/bj0530340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. FAGER E. W., ROSENBERG J. L. Phosphoglyceric acid in photosynthesis. Science. 1950 Nov 24;112(2917):617–618. doi: 10.1126/science.112.2917.617. [DOI] [PubMed] [Google Scholar]
  12. FRISELL W. R., MEECH L. A., MACKENZIE C. G. A simplified photometric analysis for serine and formaldehyde. J Biol Chem. 1954 Apr;207(2):709–716. [PubMed] [Google Scholar]
  13. GIBSON D. M., DAVISSON E. O., BACHHAWAT B. K., RAY B. R., VESTLING C. S. Rat liver lactic dehydrogenase. I. Isolation and chemical properties of the crystalline enzyme. J Biol Chem. 1953 Jul;203(1):397–409. [PubMed] [Google Scholar]
  14. GLOCK G. E., MCLEAN P. Levels of oxidized and reduced diphosphopyridine nucleotide and triphosphopyridine nucleotide in animal tissues. Biochem J. 1955 Nov;61(3):388–390. doi: 10.1042/bj0610388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. GREENBERG D. M., ICHIHARA A. Further studies on the pathway of serine formation from carbohydrate. J Biol Chem. 1957 Jan;224(1):331–340. [PubMed] [Google Scholar]
  16. HAKALA M. T., GLAID A. J., SCHWERT G. W. Lactic dehydrogenase. II. Variation of kinetic and equilibrium constants with temperature. J Biol Chem. 1956 Jul;221(1):191–209. [PubMed] [Google Scholar]
  17. HEDRICK J. L., SALLACH H. J. The metabolism of hydroxypyruvate. II. The enzymatic oxidation and decarboxylation of hydroxypyruvate. J Biol Chem. 1961 Jul;236:1872–1875. [PubMed] [Google Scholar]
  18. HEINZ F., BARTELSEN K., LAMPRECHT W. [D-glycerate dehydrogenase from liver. Contribution to serine metabolism]. Hoppe Seylers Z Physiol Chem. 1962 Nov 15;329:222–240. doi: 10.1515/bchm2.1962.329.1.222. [DOI] [PubMed] [Google Scholar]
  19. HEINZ F., LAMPRECHT W. [D-Glycerate dehydrogenase in the liver. Preliminary report]. Hoppe Seylers Z Physiol Chem. 1961 Sep 20;325:280–282. doi: 10.1515/bchm2.1961.325.1.280. [DOI] [PubMed] [Google Scholar]
  20. HOLZER H., HOLLDORF A. Isolierung von D-Glycerat-dehydrogenase, einige Eigenschaften des Enzyms und seine Verwendung zur enzymatisch-optischen Bestimmung von Hydroxypyruvat neben Pyruvat. Biochem Z. 1957;329(4):292–312. [PubMed] [Google Scholar]
  21. HORECKER B. L., SMYRNIOTIS P. Z., KLENOW H. The formation of sedoheptulose phosphate. J Biol Chem. 1953 Dec;205(2):661–682. [PubMed] [Google Scholar]
  22. ICHIHARA A., GREENBERG D. M. Studies on the purification and properties of D-glyceric acid kinase of liver. J Biol Chem. 1957 Apr;225(2):949–958. [PubMed] [Google Scholar]
  23. JAGANNATHAN V., SCHWEET R. S. Pyruvic oxidase of pigeon breast muscle. I. Purification and properties of the enzyme. J Biol Chem. 1952 May;196(2):551–562. [PubMed] [Google Scholar]
  24. KATTERMANN R., DOLD U., HOLZER H. [D-Glycerate in fructose degradation in the liver]. Biochem Z. 1961;334:218–226. [PubMed] [Google Scholar]
  25. KREBS H. A., KORNBERG H. L., BURTON K. A survey of the energy transformations in living matter. Ergeb Physiol. 1957;49:212–298. [PubMed] [Google Scholar]
  26. LABEYRIE F., NASLIN L., CURDEL A., WURMSER R. [A new determination of the redox potential of the lactate-pyruvate system]. Biochim Biophys Acta. 1960 Jul 15;41:509–515. doi: 10.1016/0006-3002(60)90049-4. [DOI] [PubMed] [Google Scholar]
  27. LAMPRECHT W., HEINZ F., DIAMANTSTEINT [Phosphorylation of D-glyceric acid to 2-phospho-D-glyceric acid with glycerate kinase in the liver. II. Identification of the reaction products by paper chromatography]. Hoppe Seylers Z Physiol Chem. 1962 Aug 6;328:204–206. doi: 10.1515/bchm2.1962.328.1.204. [DOI] [PubMed] [Google Scholar]
  28. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  29. MEISTER A. Enzymatic preparation of alpha-keto acids. J Biol Chem. 1952 May;197(1):309–317. [PubMed] [Google Scholar]
  30. MEISTER A., FRASER P. E., TICE S. V. Enzymatic desulfuration of beta-mercaptopyruvate to pyruvate. J Biol Chem. 1954 Feb;206(2):561–575. [PubMed] [Google Scholar]
  31. MEISTER A. Reduction of alpha gamma-diketo and alpha-keto acids catalyzed by muscle preparations and by crystalline lactic dehydrogenase. J Biol Chem. 1950 May;184(1):117–129. [PubMed] [Google Scholar]
  32. NEILANDS J. B. Studies on lactic dehydrogenase of heart. I. Purity, kinetics, and equilibria. J Biol Chem. 1952 Nov;199(1):373–381. [PubMed] [Google Scholar]
  33. SALLACH H. J. Formation of serine hydroxypryuvate and L-alanine. J Biol Chem. 1956 Dec;223(2):1101–1108. [PubMed] [Google Scholar]
  34. SAVAGE N. Preparation and properties of highly purified diaphorase. Biochem J. 1957 Sep;67(1):146–155. doi: 10.1042/bj0670146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. SCHWERT G. W., HAKALA M. T. Lactic dehydrogenase. I. Kinetics. Arch Biochem Biophys. 1952 Jul;38:55–65. doi: 10.1016/0003-9861(52)90009-x. [DOI] [PubMed] [Google Scholar]
  36. SMITH V. H. Removal of internally deposited plutonium. Nature. 1958 Jun 28;181(4626):1792–1793. doi: 10.1038/1811792a0. [DOI] [PubMed] [Google Scholar]
  37. STAFFORD H. A., MAGALDI A., VENNESLAND B. The enzymatic reduction of hydroxypyruvic acid to D-glyceric acid in higher plants. J Biol Chem. 1954 Apr;207(2):621–629. [PubMed] [Google Scholar]
  38. Straub F. B. Crystalline lactic dehydrogenase from heart muscle. Biochem J. 1940 Apr;34(4):483–486. doi: 10.1042/bj0340483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. TUBBS P. K., GREVILLE G. D. The oxidation of D-alpha-hydroxy acids in animal tissues. Biochem J. 1961 Oct;81:104–114. doi: 10.1042/bj0810104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. WILLIS J. E., SALLACH H. J. Activation of mammalian glycerate dehydrogenase by inorganic salts. Biochim Biophys Acta. 1962 Aug 13;62:443–444. doi: 10.1016/0006-3002(62)90282-2. [DOI] [PubMed] [Google Scholar]
  41. WILLIS J. E., SALLACH H. J. Evidence for a mammalian D-glyceric dehydrogenase. J Biol Chem. 1962 Mar;237:910–915. [PubMed] [Google Scholar]
  42. ZELITCH I. The isolation and action of crystalline glyoxylic acid reductase from tobacco leaves. J Biol Chem. 1955 Oct;216(2):553–575. [PubMed] [Google Scholar]

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