Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1970 Jan;116(1):125–134. doi: 10.1042/bj1160125

Properties of a halophil nicotinamide–adenine dinucleotide phosphate-specific isocitrate dehydrogenase. Preliminary studies of the salt relations and kinetics of the crude enzyme

D M Aitken 1, A J Wicken 1, A D Brown 1
PMCID: PMC1185331  PMID: 4391514

Abstract

The effects of chlorides on NADP-specific isocitrate dehydrogenase from Halobacterium salinarium were investigated. The enzyme is stabilized by potassium chloride and sodium chloride and this effect is discussed in relation to the Hill (1913) equation. Kinetics of the enzyme were studied within a range of concentrations of potassium chloride and sodium chloride. Apparent Michaelis constants for both substrates were affected by salt concentration, the effect being greater in sodium chloride than in potassium chloride. Minimal apparent Michaelis constants for both substrates were similar to the corresponding constants reported for yeast isocitrate dehydrogenase. Vmax. was maximal in each salt at a concentration of about 1m. The maximum was higher in sodium chloride than in potassium chloride. At salt concentrations above about 2.3m, the apparent Vmax. was lower in sodium chloride than in potassium chloride, and at salt concentrations below 0.75–1.0m, each salt behaved as a linear activator of the enzyme. Within this concentration range salt and NADP+ acted competitively; the activation by salt was overcome at finite concentrations of NADP+. At concentrations above about 1m, potassium chloride was a linear non-competitive inhibitor of the enzyme. Within the range 1.0–2.5m, sodium chloride was also a linear non-competitive inhibitor, but above 2.5m it caused more pronounced inhibition.

Full text

PDF
125

Selected References

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

  1. Aitken D. M., Brown A. D. Citrate and glyoxylate cycles in the halophil, Halobacterium salinarium. Biochim Biophys Acta. 1969 Apr 1;177(2):351–354. doi: 10.1016/0304-4165(69)90148-2. [DOI] [PubMed] [Google Scholar]
  2. BAXTER R. M. An interpretation of the effects of salts on the lactic dehydrogenase of Halobacterium salinarium. Can J Microbiol. 1959 Feb;5(1):47–57. doi: 10.1139/m59-006. [DOI] [PubMed] [Google Scholar]
  3. BAXTER R. M., GIBBONS N. E. Effects of sodium and potassium chloride on certain enzymes of Micrococcus halodenitrificans and Pseudomonas salinaria. Can J Microbiol. 1956 Oct;2(6):599–606. doi: 10.1139/m56-072. [DOI] [PubMed] [Google Scholar]
  4. BROWN A. D. ASPECTS OF BACTERIAL RESPONSE TO THE IONIC ENVIRONMENT. Bacteriol Rev. 1964 Sep;28:296–329. doi: 10.1128/br.28.3.296-329.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bernofsky C., Utter M. F. Mitochondrial isocitrate dehydrogenases from yeast. J Biol Chem. 1966 Nov 25;241(22):5461–5466. [PubMed] [Google Scholar]
  6. CHRISTIAN J. H., WALTHO J. A. Solute concentrations within cells of halophilic and non-halophilic bacteria. Biochim Biophys Acta. 1962 Dec 17;65:506–508. doi: 10.1016/0006-3002(62)90453-5. [DOI] [PubMed] [Google Scholar]
  7. CLELAND W. W. The kinetics of enzyme-catalyzed reactions with two or more substrates or products. III. Prediction of initial velocity and inhibition patterns by inspection. Biochim Biophys Acta. 1963 Feb 12;67:188–196. doi: 10.1016/0006-3002(63)91816-x. [DOI] [PubMed] [Google Scholar]
  8. HOLMES P. K., HALVORSON H. O. PROPERTIES OF A PURIFIED HALOPHILIC MALIC DEHYDROGENASE. J Bacteriol. 1965 Aug;90:316–326. doi: 10.1128/jb.90.2.316-326.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hill A. V. The Combinations of Haemoglobin with Oxygen and with Carbon Monoxide. I. Biochem J. 1913 Oct;7(5):471–480. doi: 10.1042/bj0070471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hochstein L. I., Dalton B. P. Salt specificity of a reduced nicotinamide adenine dinucleotide oxidase prepared from a halophilic bacterium. J Bacteriol. 1968 Jan;95(1):37–42. doi: 10.1128/jb.95.1.37-42.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KORNBERG A., PRICER W. E., Jr Di- and triphosphopyridine nucleotide isocitric dehydrogenases in yeast. J Biol Chem. 1951 Mar;189(1):123–136. [PubMed] [Google Scholar]
  12. 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]
  13. MOYLE J. Some properties of purified isocitric enzyme. Biochem J. 1956 Aug;63(4):552–558. doi: 10.1042/bj0630552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SANWAL B. D., ZINK M. W., STACHOW C. S. NICOTINAMIDE ADENINE DINUCLEOTIDE-SPECIFIC ISOCITRIC DEHYDROGENASE. A POSSIBLE REGULATORY PROTEIN. J Biol Chem. 1964 May;239:1597–1603. [PubMed] [Google Scholar]
  15. SEHGAL S. N., GIBBONS N. E. Effect of some metal ions on the growth of Halobacterium cutirubrum. Can J Microbiol. 1960 Apr;6:165–169. doi: 10.1139/m60-018. [DOI] [PubMed] [Google Scholar]
  16. VICKERY H. B. A suggested new nomenclature for the isomers of isocitric acid. J Biol Chem. 1962 Jun;237:1739–1741. [PubMed] [Google Scholar]

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

RESOURCES