Purification and characterization of rat adrenal 3 beta-hydroxysteroid dehydrogenase with steroid 5-ene-4-ene-isomerase.
After solubilization of rat adrenal microsomes with sodium cholate, 3 beta-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: 3 beta-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. Either NAD+ or NADH was required for demonstration of steroid isomerase activity. Treatment of the enzyme with 5'-p-fluorosulfonylbenzoyladenosine, an affinity labeling reagent for NAD+-dependent enzyme, diminished both the enzyme activities.
D015096:3-Hydroxysteroid Dehydrogenases / Q000032:analysis / Q000302:isolation & purification*; D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D003687:Dehydroepiandrosterone / Q000378:metabolism; D007535:Isomerases / Q000302:isolation & purification*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D009243:NAD / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013251:Steroid Isomerases / Q000032:analysis / Q000302:isolation & purification*; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000502:physiology
D013438:Sulfhydryl Compounds; D009243:NAD; D003687:Dehydroepiandrosterone; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D015096:3-Hydroxysteroid Dehydrogenases; D007535:Isomerases; D013251:Steroid Isomerases; D000241:Adenosine