title: Rat liver microsomal 3 alpha-hydroxysteroid dehydrogenase and dihydrodiol dehydrogenase: solubilization, separation and partial purification.
abstract: Rat liver microsomes contain 3 alpha-hydroxysteroid dehydrogenase (HSD) (EC 1.1.1.50) and dihydrodiol dehydrogenase (DHD) (EC 1.3.1.20) activities. The two enzyme activities were solubilized by 10% Triton X-100 or 0.4% sodium deoxycholate. Unlike the cytosolic enzyme (Penning & Talalay (1983) Proc. Natl. Acad. Sci. U.S.A., 80, 4505), the microsomal HSD and DHD activities were not inhibited by indomethacin. Chromatography of the microsomal Triton X-100 extract on Affigel Blue and then on Phenyl-Sepharose gave an HSD preparation containing no detectable (less than 3 - 5%) DHD activity, whereas chromatography of the deoxycholate extract on Phenyl-Sepharose provided a DHD preparation that lacked measurable HSD activity. These results are in sharp contrast to the cytosolic enzyme where both HSD and DHD activities could be copurified to homogeneity (Penning et al. (1984) Biochem. J. 222, 601).
mesh_terms: D015096:3-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000818:Animals; D003840:Deoxycholic Acid; D006863:Hydrogen-Ion Concentration; D007213:Indomethacin / Q000494:pharmacology; D007425:Intracellular Membranes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; D051381:Rats; D012995:Solubility
chemical_list: D003840:Deoxycholic Acid; D009243:NAD; D010088:Oxidoreductases; D015096:3-Hydroxysteroid Dehydrogenases; D000429:Alcohol Oxidoreductases; C023125:dihydrodiol dehydrogenases; D044925:Oxidoreductases Acting on CH-CH Group Donors; C116788:cis-1,2-dihydro-1,2-dihydroxynaphthalene dehydrogenase; D007213:Indomethacin