title: Kinetic and stereochemical studies on reaction mechanism of mouse liver 17 beta-hydroxysteroid dehydrogenases.
abstract: The kinetic mechanism of two major monomeric 17 beta-hydroxysteroid dehydrogenases from mouse liver cytosol was studied at pH 7 in both directions with NADP(H) and three steroid substrates: testosterone, 5 beta-androstane-3 alpha, 17 17 beta-diol, and estradiol-17 beta. In each case the reaction mechanism of the two enzymes was sequential, and inhibition patterns by-products and dead-end inhibitors were consisted with an ordered bi bi mechanism with the coenzyme binding to the free enzyme, although there was difference in affinity and maximum velocity for the steroidal substrates between the two enzymes. Binding studies of the coenzyme and substrate indicate the binding of coenzyme to the free enzyme, in which 1 mol of NADPH binds to 1 mol of each monomeric enzyme. The 4-pro-R-hydrogen atom of NADPH was transferred to the alpha-face of the steroid molecule by the two enzymes.
authors.affiliation: Department of Biochemistry, Gifu Pharmaceutical University.
authors.forename: A
authors.initials: A
authors.lastname: Hara
authors.forename: T
authors.initials: T
authors.lastname: Nakayama
authors.forename: M
authors.initials: M
authors.lastname: Nakagawa
authors.forename: Y
authors.initials: Y
authors.lastname: Inoue
authors.forename: H
authors.initials: H
authors.lastname: Tanabe
authors.forename: H
authors.initials: H
authors.lastname: Sawada
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D015113:Androstane-3,17-diol / Q000378:metabolism; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004958:Estradiol / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D051379:Mice; D008968:Molecular Conformation; D009249:NADP / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D015113:Androstane-3,17-diol; D013739:Testosterone; D004958:Estradiol; D009249:NADP; D015067:17-Hydroxysteroid Dehydrogenases; C032690:3-alpha-(17-beta)-hydroxysteroid dehydrogenase (NAD(+))