title: 5-ene-3beta-hydroxysteroid dehydrogenase of human and bovine adrenocortical endoplasmic reticulum: solubilization and fractionation.
abstract: Protein moieties of various molecular sizes and possessing 5-ene-3beta-hydroxysteroid dehydrogenase activity have been successfully solubilized from the microsomal membranes of both bovine and human adrenal glands using a combination of Triton X-100 and sonication. These moieties have been studied by gel filtration, sucrose density gradient centrifugation and isoelectric focusing, and were shown to possess a minimum molecular weight of about 118 000, with an isoelectric point between 7-2 and 7-4. The molecular weight was dependent upon the concentration of Triton X-100 used during fractionation. No separation of dehydrogenase activities toward the three steroid substrates, pregnenolone, 17alpha-hydroxy-pregnenolone and dehydroisoandrosterone, was observed. Changes in the relative activities for the steroid substrates during fractionation were observed, but have been attributed to the formation of allotypes rather than to the existence of separate dehydrogenases with restricted substrated specificity.
mesh_terms: D006907:17-alpha-Hydroxypregnenolone / Q000494:pharmacology; D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D003687:Dehydroepiandrosterone / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D006801:Humans; D006913:Hydroxysteroid Dehydrogenases / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011284:Pregnenolone / Q000494:pharmacology
chemical_list: D006907:17-alpha-Hydroxypregnenolone; D003687:Dehydroepiandrosterone; D011284:Pregnenolone; D006913:Hydroxysteroid Dehydrogenases