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Showing papers on "Hydroxysteroid dehydrogenase published in 1989"


Journal ArticleDOI
TL;DR: 1 H-NMR and EI-MS of the nicotinamide moiety after enzymatic oxidation of deuterated NAD(P)H with dehydrocholic acid as substrate showed that both dehydrogenases are B-sterospecific.

50 citations


30 Jun 1989
TL;DR: The results indicate that the major form of dihydrodiol dehydrogenase is identical to 17 beta-hydroxysteroid dehydrogen enzyme and the minor enzyme form to aldehyde reductase, which reduced various aldehydes well and was specifically inhibited by barbiturates and sorbinil.
Abstract: A major and a minor form of dihydrodiol dehydrogenase were co-purified with 17 beta-hydroxysteroid dehydrogenase and aldehyde reductase, respectively, to apparent homogeneity from liver cytosol of male ddY mice. The activities of dihydrodiol dehydrogenase and testosterone dehydrogenase or aldehyde reductase of the two enzyme forms comigrated electrophoretically. The major form of the enzyme oxidized 17 beta-hydroxysteroids and nonsteroidal alicyclic alcohols and reduced 17-ketosteroids and various synthetic carbonyl compounds, showing higher affinity for steroids than for xenobiotics. The activity of this enzyme form toward benzene dihydrodiol and testosterone exhibited identical thermostability and susceptibility to inhibition by quercitrin, SH-reagents, nonsteroidal estrogens and anti-inflammatory agents. On the other hand, the minor form of the enzyme, which oxidized benzene dihydrodiol but not 17 beta-hydroxysteroids, also reduced various aldehydes well and was specifically inhibited by barbiturates and sorbinil. These results indicate that the major form of dihydrodiol dehydrogenase is identical to 17 beta-hydroxysteroid dehydrogenase and the minor enzyme form to aldehyde reductase.

34 citations


Journal ArticleDOI
TL;DR: It is proposed that the steroid recognition site on 17β-OH-steroid dehydrogenase evolved from an ancestral recognition site for polyols such as ribitol and glucitol-6-phosphate.
Abstract: The amino acid sequence of human placental 17β-hydroxysteroid dehydrogenase (17β-OH-steroid dehydrogenase) was found to be similar to that of the NodG protein of Rhizobium meliloti. The computer-based comparison score is 11.5 SD higher than that obtained with 2500 comparisons of randomized sequences of these proteins. The probability of getting such a score by chance is 6 × 10−31. 17β-OH-steroid dehydrogenase is also similar to Klebsiella aerogenes ribitol dehydrogenase and Escherichia coli glucitol-6-phosphate dehydrogenase. We propose that the steroid recognition site on 17β-OH-steroid dehydrogenase evolved from an ancestral recognition site for polyols such as ribitol and glucitol-6-phosphate.

32 citations


Journal ArticleDOI
TL;DR: A human fecal isolate, characterized by morphological, physiological and biochemical data as a strain of Peptostreptococcus roductus, was shown to contain NAD-dependent 3α- and 3β-hydroxysteroid dehydrogenases and a NADP-dependent 7β-Hydroxysteroids dehydrogenase, which could be demonstrated in crude extracts and in membrane fractions.

32 citations



Journal ArticleDOI
TL;DR: Dihydrodiol dehydrogenase activity was detected in the cytosol of various mouse tissues, among which kidney exhibited high specific activity comparable to the value for liver as mentioned in this paper.
Abstract: Dihydrodiol dehydrogenase activity was detected in the cytosol of various mouse tissues, among which kidney exhibited high specific activity comparable to the value for liver. The enzyme activity in the kidney cytosol was resolved into one major and three minor peaks by Q-Sepharose chromatography: one minor form cross-reacted immunologically with hepatic 3 alpha-hydroxysteroid dehydrogenase and another with aldehyde reductase. The other minor form was partially purified and the major form was purified to homogeneity. These two forms, although different in their charges, were monomeric proteins with the same molecular weight of 39,000 and had similar catalytic properties. They oxidized cis-benzene dihydrodiol and alicyclic alcohols as well as trans-dihydrodiols of benzene and naphthalene in the presence of NADP+ or NAD+, and reduced several xenobiotic aldehydes and ketones with NAD(P)H as a cofactor. The enzymes also catalyzed the oxidation of 3 alpha-hydroxysteroids and epitestosterone, and the reduction of 3- and 17-ketosteroids, showing much lower Km values (10(-7)-10(-6) M) for the steroids than for the xenobiotic alcohols. The results of mixed substrate experiments, heat stability, and activity staining on polyacrylamide gel electrophoresis suggested that, in the two enzymes, both dihydrodiol dehydrogenase and 3(17)alpha-hydroxysteroid dehydrogenase activities reside on a single enzyme protein. Thus, dihydrodiol dehydrogenase existed in four forms in mouse kidney cytosol, and the two forms distinct from the hepatic enzymes may be identical to 3(17)alpha-hydroxysteroid dehydrogenases.

22 citations


Journal ArticleDOI
TL;DR: Results on the kinetics of 7α-hydroxysteroid dehydrogenase 7 α-HSDH showed that this enzyme could oxidize all bile acids having an –OH group at the C-7 position and a constant increase in the enzyme activity with increase in enzyme-protein concentration.
Abstract: Results on the kinetics of 7α-hydroxysteroid dehydrogenase 7α-HSDH showed that this enzyme could oxidize all bile acids having an –OH group at the C-7 position. Lineweaver-Burk plots showed Michael...

21 citations


Journal ArticleDOI
TL;DR: The results strongly suggest that indanol dehydrogenase acts as a 3(20)alpha-hydroxysteroid dehydrogen enzyme in the metabolism of certain steroid hormones and bile acids.
Abstract: Homogeneous indanol dehydrogenase from monkey liver catalyzed the reversible conversion of 3 alpha- or 20 alpha-hydroxy groups of several bile acids and 5 beta-pregnanes to the corresponding 3- or 20-ketosteroids. The kcat values for the steroids determined at pH 7.4 were low, but the kcat/Km values for the 3-ketosteroids were comparable to or exceeded those for 1-indanol and xenobiotic carbonyl substrates. The enzyme transferred the 4-pro-R-hydrogen atom of NADPH to the 3 beta- or 20 beta-face of the ketosteroid substrate. Competitive inhibition of the hydroxysteroid dehydrogenase activity of the enzyme by medroxyprogesterone acetate, hexestrol, and 1,10-phenanthroline suggests that both 1-indanol and hydroxysteroid are oxidized at the same active site on the enzyme. The specific inhibitor of the enzyme, 1,10-phenanthroline, suppressed the 3 alpha-hydroxysteroid dehydrogenase activity in the crude extract of monkey liver by 50%. The results strongly suggest that indanol dehydrogenase acts as a 3(20)alpha-hydroxysteroid dehydrogenase in the metabolism of certain steroid hormones and bile acids.

11 citations


Journal ArticleDOI
TL;DR: The basic for undertaking the present investigation was provided by results of preliminary experiments showing the presence of a nonreceptor protein, intensively binding 3H-estradiol, in the soluble fraction of rabbit liver, which possesses oxidoreductase activity relative to androgens and gestagens.
Abstract: One of the mechanisms of regional regulation of steroid hormone reception and metabolism may be reversible interaction of these hormones with specific intracellular nonreceptor proteins, or steromodulins [3, 5, 7]. This group of substances includes proteins similar or identical to blood transport proteins, and specialized, tissue-specific proteins of the special estrogen-binding protein (SEBP) type of rat liver [I, 12, 13]. The basic for undertaking the present investigation was provided by results of preliminary experiments showing the presence of a nonreceptor protein, intensively binding 3H-estradiol, in the soluble fraction of rabbit liver. It was found in the course of the work that this protein possesses oxidoreductase activity relative to androgens and gestagens.