Abstract

After solubilization of rat adrenal microsomes with sodium cholate, 3β-hydroxysteroid dehydrogenase with steroid 5-ene-4-ene isomerase (abbreviated as steroid isomerase) activity was purified to a homogeneous state. The following characteristics of the enzyme were obtained: (1) 3β-Hydroxysteroid dehydrogenase together with steroid isomerase was detected as a single protein band in SDS-polyacrylamide gel electrophoresis, where its mol. wt was estimated as 46,500. (2) Either NAD + or NADH was required for demonstration of steroid isomerase activity. (3) Treatment of the enzyme with 5'- p-fluorosulfonylbenzoyladenosine, an affinity labeling reagent for NAD +-dependent enzyme, diminished both the enzyme activities.

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