Purification of yeast isocitrate dehydrogenase.
The NAD-linked isocitrate dehydrogenase from baker's yeast was purified to homogeneity (as judged by gel filtration and polyacrylamide-gel electrophoresis) with an overall yield of 50% by using dilute solutions of the allosteric effector (AMP) to elute the enzyme specifically from CM-cellulose. This method preserves the allosteric properties of the crude enzyme. Although the pure enzyme shows only a single band on electrophoresis in the presence of sodium dodecyl sulphate, two types of subunit are observed in 8m-urea. The isoelectric point of the enzyme rises during purification, and this may reflect the partial loss of an additional low-molecular-weight component. Values are included for the amino acid composition and extinction coefficients of the pure enzyme.
D000249:Adenosine Monophosphate; D000596:Amino Acids / Q000032:analysis; D002498:Centrifugation; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005615:Freezing; D006863:Hydrogen-Ion Concentration; D007521:Isocitrate Dehydrogenase / Q000302:isolation & purification*; D007700:Kinetics; D008722:Methods; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D000596:Amino Acids; D000249:Adenosine Monophosphate; D007521:Isocitrate Dehydrogenase