1887

Abstract

Malate dehydrogenase (MDH; EC 1.1.1.37) from was purified and characterized. MDH activity from whole cells was purified 275-fold. The mitochondrial cytoplasmic MDH present co-purified through three ion exchange and affinity chromatography steps. The isoenzymes were barely separable by either disc gel electrophoresis or isoelectric focusing. The purified preparation containing both isoenzymes had a single pH optimum (9·3–9·5) and an apparent molecular weight of 70000. It exhibited linear kinetics and responded to known inhibitors of MDH, i.e. thyroxine and hydroxymalonate. Michaelis and dissociation constants obtained with this preparation were similar to those obtained with a 10-fold purified mitochondrial MDH.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-8-1767
1982-08-01
2024-12-10
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/8/mic-128-8-1767.html?itemId=/content/journal/micro/10.1099/00221287-128-8-1767&mimeType=html&fmt=ahah

References

  1. Banaszak L. J., Bradshaw R. A. 1975; Malate dehydrogenases. In The Enzymes, 3rd. 11 pp. 369–396 Boyer P. D. Edited by New York:: Academic Press.;
    [Google Scholar]
  2. Cleland W. W. 1963; The kinetics of enzyme-catalyzed reactions with two or more substrates or products. Biochimica et biophysica acta 67:104–137
    [Google Scholar]
  3. Flury V., Heer B., Fiechter A. 1974; Regulatory and physiochemical properties of two isozymes of malate dehydrogenase from Schizo-saccharomyces pombe. . Biochimica et biophysica acta 341:465–483
    [Google Scholar]
  4. Hagele E., Neefe J., Mecke D. 1978; The malate dehydrogenase isozymes of Saccharomyces cerevisiae. . European Journal of Biochemistry 83:67–76
    [Google Scholar]
  5. Heyde E., Ainsworth S. 1968; Kinetic studies on the mechanism of the malate dehydrogenase reaction. Journal of Biological Chemistry 243:2413–2423
    [Google Scholar]
  6. Hohorst H. J. 1965; Determination of malate with malic dehydrogenase and DPN. In Methods of Enzymatic Analysis, 1st. pp. 328–332 Bergmeyer H. U. Edited by New York:: Academic Press.;
    [Google Scholar]
  7. Kelly P. J., Kelleher J. K., Wright B. E. 1979a; Tricarboxylic acid cycle in Dictyostelium discoideum. Metabolite concentrations, oxygen uptake and l4C-labelled amino acid labelling patterns. Biochemical Journal 184:581–588
    [Google Scholar]
  8. Kelly P. J., Kelleher J. K., Wright B. E. 1979b; Tricarboxylic acid cycle in Dictyostelium discoideum. A model of the cycle at preculmination and aggregation. Biochemical Journal 184:589–597
    [Google Scholar]
  9. Kitto G. B., Kaplan N. O. 1966; Purification and properties of chicken heart mitochondrial and supernatant malic dehydrogenases. Biochemistry 5:3966–3980
    [Google Scholar]
  10. Komuniecki P. R., DeToma F. J., Lawrence M. H., didomenico L. 1980; ADP phosphorylation and glutamate oxidation in mitochondria isolated from Dictyostelium discoideum amoebae. Biochemical and Biophysical Research Communications 96:1017–1023
    [Google Scholar]
  11. Krebs H. A., Gascoyne J., Notton B. N. 1967; Generation of extramitochondrial reducing power in gluconeogenesis. Biochemical Journal 102:275–282
    [Google Scholar]
  12. Kuan K. N., Jones G. L., Vestling C. S. 1979; Rapid preparation of mitochondrial malate dehydrogenase from rat liver and heart. Biochemistry 18:4366–4373
    [Google Scholar]
  13. Marshall R., Sargent D., Wright B. E. 1970; Glycogen turnover in Dictyostelium discoideum. . Biochemistry 9:3087–3094
    [Google Scholar]
  14. Murphy W. H., Kitto G. B., Everse J., Kaplan N. O. 1967; Malate dehydrogenase. I. A survey of molecular size measured by gel filtration. Biochemistry 6:603–610
    [Google Scholar]
  15. Raval D. N., Wolfe R. G. 1962; Malic dehydrogenase. II. Kinetic studies of the reaction mechanism. Biochemistry 1:263–269
    [Google Scholar]
  16. Teague W. M., Henney H. R. Jr 1973; Purification and properties of cytoplasmic and mitochondrial malate dehydrogenases of Physarum polycephalum. . Journal of Bacteriology 116:673–684
    [Google Scholar]
  17. Thompson S. T., Cass K. H., Stellwagen E. 1975; Blue dextran sepharose: an affinity column for the dinucleotide folds in proteins. Proceedings of the National Academy of Sciences of the United States of America 72:669–672
    [Google Scholar]
  18. Wolff J., Wolff E. C. 1957; The effect of thyroxine on isolated dehydrogenases. Biochimica et biophysica acta 26:387–396
    [Google Scholar]
  19. Wright B. E. 1964; Biochemistry of Acrasiales. In Biochemistry and Physiology of the Protozoa 3 pp. 341–381 Hutner S. H. Edited by New York:: Academic Press.;
    [Google Scholar]
  20. Wright B. E. 1981; A transition model of the citric acid cycle in Dictyostelium. . Abstracts of the Annual Meeting, American Society for Microbiology abstract 1144 p. 111
    [Google Scholar]
/content/journal/micro/10.1099/00221287-128-8-1767
Loading
/content/journal/micro/10.1099/00221287-128-8-1767
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error