No abstract

1. One mitochondrial and one cytoplasmic malate dehydrogenase isoenzyme could be purified from acetate grown cells of the yeast Saccharomyces cerevisiae. 2. The purification procedure uses chromatography on dextran blue columns as an essential step for enrichment, and reverse ammonium sulfate chromatography on celite for isoenzyme separation. 3. The homogeneity of the preparations was established by gel electrophoreses in the presence of sodium dodecylsulfate and by a sedimentation run in the analytical ultracentrifuge. 4. Both enzymes are dimers with a molecular weight of 75 000 for the cytoplasmic and of 68 000 for the mitochondrial enzyme. 5. Amino acid analysis and peptide mapping showed that both enzymes are closely related, but genetically different (true isoenzymes). 6. The cytoplasmic enzyme shows electrophoretic splitting. This is most likely due to post‐translational deamination in vivo. 7. Antibodies to both isoenzymes could be obtained in rabbits. The antisera to cytoplasmic malate dehydrogenase were specific for this enzyme. Antisera to mitochondrial malate dehydrogenase react with both isoenzymes. Neither type of antisera precipitated an inactive protein after the glucose‐dependent inactivation of cytoplasmic malate dehydrogenase in vivo.

The cytoplamic malate dehydrogenase of Sacchoromyces cerevisiae was radioactively labeled during its synthesis on a glucose‐free derepression medium. After purification a sensitive radio‐immunoassay for this enzyme could be developed. The assay showed that after the physiological, glucose‐dependent ‘catabolite inactivation’ of cytoplasmic malate dehydorgenase an inactive enzyme protein is immunologically not detchtable. Together with the irreversiblity of this reaction in vivo this finding strongly suggests a proteolytic mechanism of enzyme inactivation. For this process the term ‘catabolite degradation’ is used.

The cytoplasmic malate dehydrogenase in the yeast Saccharomyces cerevisiae is known to be inactivated by a glucose dependent process. In this paper it is shown that in vivo effectors of the glucose metabolism (arsenate, iodoacetate, acetaldhyde) inhibit the inactivation or change the inactivation kinetics. In vitro it was possible to inactivate the malate dehydrogenase by addition of the glucose metabolite glyceraldehyde 3-phosphate. The physiological relevance of this modification and the effect of malate dehydrogenase inactivation on the glyoxylate cycle in yeast is discussed.