Adult-male-specific S-warfarin (11S-OH) and progesterone (20 beta-OH) keto-reductases in rat hepatic microsomes are not identical.
Adult-male-specific reductase activities in rat hepatic microsomes use NADPH to reduce S-warfarin and progesterone to their 11S-OH and 20 beta-OH products, respectively (Apanovitch et al. (1992) Biochem. Biophys. Res. Commun. 184, 338-346). When microsomes were treated with increasing concentrations of detergent, S-warfarin (11S-OH) reductase (SW(11S)R) activity was subject to monophasic activation by Triton X-100, monophasic inhibition by sodium cholate, and, activation followed by inhibition with either CHAPS or dodecyl-beta-D-maltoside. A non-dialyzable, heat-sensitive factor in rat and rabbit sera activates microsomal SW(11S)R activity six- to eight-fold. Similar detergent inhibitions but no detergent or serum activations were observed for progesterone (20 beta-OH) reductase (P(20 beta)R) activity. A significant amount of SW(11S)R activity was lost during purification regardless of whether the detergent used for solubilization was activating or inhibiting. Octyl-Sepharose, hydroxyapatite, DEAE-cellulose and carboxymethyl matrices were used to partially purify SW(11S)R. P(20 beta)R activity co-purified with SW(11S)R and the most purified fraction contained two major and several minor polypeptides. Partially purified SW(11S)R is activated by detergents, serum, and salt. These and previous results indicate that SW(11S)R and P(20 beta)R are not identical even though they are both adult male-specific, integral membrane proteins apparently having their active sites exposed on the cytoplasmic surface of the endoplasmic reticulum.
Apanovitch
D
D
Department of Chemistry, Kent State University, OH 44242, USA.
Walz
F G
FG
D015089:20-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001769:Blood; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D006908:Hydroxyprogesterones / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D008970:Molecular Weight; D011817:Rabbits; D051381:Rats; D011916:Rats, Inbred F344; D014859:Warfarin / Q000378:metabolism
D065607:Cytochrome P-450 Enzyme Inhibitors; D003902:Detergents; D006908:Hydroxyprogesterones; D014859:Warfarin; D003577:Cytochrome P-450 Enzyme System; D000429:Alcohol Oxidoreductases; D015089:20-Hydroxysteroid Dehydrogenases; C074128:S-warfarin keto reductase; C074127:progesterone 20-keto-reductase