Affinity chromatography of yeast nicotinamide-adenine dinucleotide-specific isocitrate dehydrogenase on immobilized nicotinamide-adenine dinucleotide. Effects of ligands.
The method of affinity chromatography has been used for studying the effects of some ligands of yeast NAD-specific isocitrate dehydrogenase on the affinity of the enzyme for NAD+ immobilized on Sepharose 4B. In absence of ligands, the enzyme is eluted from NAD+-Sepharose columns by 0.1 M phosphate buffer, pH 7.6, in a highly purified form. The elution of enzyme is accelerated by NAD+ and, more effectively, by AMP; and retarded by isocitrate and citrate. The elution patterns show a rather irregular shape, probably due to the occurrence of aggregation processes of the enzyme protein.
D000249:Adenosine Monophosphate; D002846:Chromatography, Affinity; D002951:Citrates; D007521:Isocitrate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D007523:Isocitrates; D046911:Macromolecular Substances; D009243:NAD; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D002951:Citrates; D007523:Isocitrates; D046911:Macromolecular Substances; D009243:NAD; D000249:Adenosine Monophosphate; D007521:Isocitrate Dehydrogenase