Purification and properties of testicular 3 beta-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase.
Through the treatment of rat testicular microsomes with sodium cholate, 3 beta-hydroxy-5-ene-steroid dehydrogenase and 5-ene-4-ene isomerase (abbreviated as the 3 beta-hydroxysteroid dehydrogenase and isomerase, respectively) were solubilized, and then purified by DEAE and hydroxylapatite column chromatographies. The findings were as follows: With this purification procedure, the 3 beta-hydroxysteroid dehydrogenase activity could not be separated from the isomerase. For 3-oxo-4-ene-steroid formation from 3 beta-hydroxy-5-ene-steroids, NAD+ was required as a cofactor. While the 3 beta-hydroxysteroid dehydrogenase required NAD+, the isomerase also required NAD+ or its reduced form, in contrast to the microbial enzyme. On treatment of the purified enzyme with 5'-p-fluorosulfonyl-benzoyladenosine (FSBA), both enzyme activities were markedly reduced. The enzyme, affinity labeled with [adenine-8-14C]FSBA, showed a mol. wt of 46.8 K. During 4-androstenedione production from DHA, 5-androstenedione was detected as an intermediate.
D015096:3-Hydroxysteroid Dehydrogenases / Q000302:isolation & purification*; D000241:Adenosine / Q000031:analogs & derivatives; D000345:Affinity Labels; D000735:Androstenedione / Q000096:biosynthesis; D000818:Animals; D003067:Coenzymes / Q000378:metabolism; D003687:Dehydroepiandrosterone / Q000378:metabolism; D007535:Isomerases / Q000302:isolation & purification*; D008297:Male; D008970:Molecular Weight; D051381:Rats; D012995:Solubility; D013251:Steroid Isomerases / Q000302:isolation & purification*; D013737:Testis / Q000201:enzymology*
D000345:Affinity Labels; D003067:Coenzymes; D000735:Androstenedione; D003687:Dehydroepiandrosterone; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D015096:3-Hydroxysteroid Dehydrogenases; D007535:Isomerases; D013251:Steroid Isomerases; C022660:steroid delta-isomerase; D000241:Adenosine