Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1968 Aug;108(5):793–796. doi: 10.1042/bj1080793

The resolution of some steps of the reactions of lactate dehydrogenase with its substrates

H D'A Heck 1, C H McMurray 1, H Gutfreund 1
PMCID: PMC1198886  PMID: 4299820

Abstract

1. The reaction of pig heart lactate dehydrogenase (EC 1.1.1.27) with NAD+ and lactate to form pyruvate and NADH was followed by rapid spectrophotometric methods. The distinct spectrum of enzyme-bound NADH permits the measurement of the rate of dissociation of this compound. 2. The reduction of the first mole equivalent of NAD+ per mole of enzyme sites can also be observed, and is much more rapid than the steady-state rate of NADH production. 3. At pH8 the dissociation of the enzyme–NADH complex is rate-determining for the steady-state oxidation of lactate. At lower pH some other step after the interconversion of the ternary complex and before the dissociation of NADH is rate-determining. Other evidence for a compulsory-order mechanism is provided.

Full text

PDF
793

Selected References

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

  1. Barman T. E., Gutfreund H. Optical and chemical identification of kinetic steps in trypsin- and chymotrypsin-catalysed reactions. Biochem J. 1966 Nov;101(2):411–416. doi: 10.1042/bj1010411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CHANCE B., NEILANDS J. B. Studies on lactic dehydrogenase of heart. II. A compound of lactic dehydrogenase and reduced pyridine nucleotide. J Biol Chem. 1952 Nov;199(1):383–387. [PubMed] [Google Scholar]
  3. CZERLINSKI G. H., SCHRECK G. FLUORESCENCE DETECTION OF THE CHEMICAL RELAXATION OF THE REACTION OF LACTATE DEHYDROGENASE WITH REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE. J Biol Chem. 1964 Mar;239:913–921. [PubMed] [Google Scholar]
  4. Geraci G., Gibson Q. H. The reaction of liver alcohol dehydrogenase with reduced diphosphopyridine nucleotide. J Biol Chem. 1967 Sep 25;242(18):4275–4278. [PubMed] [Google Scholar]
  5. Gutfreund H., Cantwell R., McMurray C. H., Criddle R. S., Hathaway G. The kinetics of the reversible inhibition of heart lactate dehydrogenase through the formation of the enzyme-oxidized nicotinamide-adenine dinucleotide-pyruvate compounds. Biochem J. 1968 Feb;106(3):683–687. doi: 10.1042/bj1060683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Holbrook J. J. The importance of SH-groups for enzymic activity. V. The coenzyme-binding capacity of pig heart lactate dehydrogenase, isozyme I, after inhibition by various maleinimides. Biochem Z. 1966 Mar 28;344(2):141–152. [PubMed] [Google Scholar]
  7. Iwatsubo M., Pantaloni D. Régulation de l'activité de la glutamate déshydrogènase par les effecteurs GTP et ADP: étude par "stopped flow". Bull Soc Chim Biol (Paris) 1967 Dec 18;49(11):1563–1572. [PubMed] [Google Scholar]
  8. JECKEL D., PFLEIDERER G., WIELAND T. Uber die Einwirkung von Sulfit auf einige DPN hydrierende Enzyme. Biochem Z. 1956;328(3):187–194. [PubMed] [Google Scholar]
  9. Schwert G. W., Miller B. R., Peanasky R. J. Lactic dehydrogenase. X. A re-evaluation of the effects of pH upon the kinetics of the reaction. J Biol Chem. 1967 Jul 25;242(14):3245–3252. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES