Purification and subunit structure of nicotinamide adenine dinucleotide specific isocitrate dehydrogenase from Neurospora crassa

Abstract
N. crassa NAD specific isocitrate dehydrogenase was purified to homogeneity by the criteria of disc gel electrophoresis and sedimentation equilibrium. Purification of the enzyme is facilitated by the presence of phenylmethanesulfonyl fluoride and by the use of a ribose-linked AMP affinity column. The enzyme appears to be composed of nonidentical subunits of MW 42,800 and 38,300 as estimated by polyacrylamide gel electrophoresis in 0.1% sodium dodecyl sulfate. The intensity of each band and the native MW showed that the enzyme is composed of either 6 or 8 subunits, 3 or 4 of each type, respectively. The availability of pure enzyme will allow clarification of the structure of the enzyme by ligand binding studies.

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