Steroid specificity of human placental 5-ene-3 beta-hydroxysteroid oxidoreductase.
The properties of 5-ene-3 beta hydroxysteroid oxidoreductase (3 beta-HSD) from human placental homogenates were studied in vitro. The apparent Michaelis constants for 3 beta-HSD with the substrates pregnenolone (delta 5P) and dehydroepiandrosterone (DHA) were 170 nM and nM respectively. The optimal pH for both these substrates was between 10 and 12. With NAD as the substrate, the Km for the pregnenolone was 20 microM and for DHA, 17 microM. The activity of 3 beta-HSD was inhibited by various steroids. Competitive inhibitors (pregnenolone substrate) included: ethynylestradiol (inhibition constant Ki=7.3 nM), DHA (Ki=46 nM), estradiol-17 beta (Ki=46 nM), cholesterol (Ki=0.68 microM) and 16 alpha-hydroxydehydroepiandrosterone (16 alphaOHDHA) (Ki=2.2 microM). When the substrate was DHA, competitive inhibition occurred with the following steroids: ethynylestradiol (Ki=6.4 nM), estradiol-17 beta (Ki=69 nM), pregnenolone (Ki=91 nM), cholesterol (Ki=1.3 microM) and 16 alphaOHDHA (Ki=1.9 microM). 4-Ene-3-ketosteroids such as androstenedione, progesterone (delta 4P), norethindrone and chlormadinone acetate acted as noncompetitive inhibitors towards both substrates.
D015096:3-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003687:Dehydroepiandrosterone / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006914:Hydroxysteroids / Q000494:pharmacology; D007664:Ketosteroids / Q000494:pharmacology; D009243:NAD / Q000494:pharmacology; D010920:Placenta / Q000201:enzymology*; D011247:Pregnancy; D011284:Pregnenolone / Q000378:metabolism; D013379:Substrate Specificity
D004967:Estrogens; D006914:Hydroxysteroids; D007664:Ketosteroids; D009243:NAD; D003687:Dehydroepiandrosterone; D011284:Pregnenolone; D015096:3-Hydroxysteroid Dehydrogenases