Bovine adrenal 3beta-hydroxysteroid dehydrogenase (E.C. 1.1.1. 145)/5-ene-4-ene isomerase (E.C. 5.3.3.1): characterization and its inhibition by isoflavones.
The isoflavones daidzein, genistein, biochanin A and formononetin inhibit potently and preferentially the gamma-isozymes of mammalian alcohol dehydrogenase (gammagamma-ADH), the only ADH isozyme that catalyzes the oxidation of 3beta-hydroxysteroids. Based on these results, we proposed that these isoflavones might also act on other enzymes involved in 3beta-hydroxysteroid metabolism. Recently, we showed that they indeed are potent inhibitors of a bacterial beta-hydroxysteroid dehydrogenase (beta-HSD). To extend this finding to the mammalian systems, we hereby purified, characterized and studied the effects of isoflavones and structurally related compounds on, a bovine adrenal 3beta-hydroxysteroid dehydrogenase (3beta-HSD). This enzyme catalyzes the oxidation of 3beta-hydroxysteroids but not 3alpha-, 11beta- or 17beta-hydroxysteroids. The same enzyme also catalyzes 5-ene-4-ene isomerization, converting 5-pregnen 3, 20-dione to progesterone. The K(m) values of its dehydrogenase activity determined for a list of 3beta-hydroxysteroid substrates are similar (1 to 2 microM) and that of its isomerase activity, determined with 5-pregnen 3, 20-dione as a substrate, is 10 microM. The k(cat) value determined for its isomerase activity (18.2 min(-1)) is also higher than that for its dehydrogenase activity (1.4-2.4 min(-1)). A survey of more than 30 isoflavones and structurally related compounds revealed that daidzein, genistein, biochanin A and formononetin inhibit both the dehydrogenase and isomerase activity of this enzyme. Inhibition is potent and concentration dependent. IC(50) values determined for these compounds range from 0.4 to 11 microM, within the plasma and urine concentration ranges of daidzein and genistein of individuals on vegetarian diet or semi-vegetarian diet. These results suggest that dietary isoflavones may exert their biological effects by inhibiting the action of 3beta-HSD, a key enzyme of neurosteroid and/or steroid hormone biosynthesis.
Wong
C K
CK
Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong. ck-wong@cuhk.edu.hk
Keung
W M
WM
D000311:Adrenal Glands / Q000201:enzymology*; D000818:Animals; D001192:Arylsulfatases / Q000378:metabolism; D002417:Cattle; D003374:Coumarins / Q000494:pharmacology; D006224:Cricetinae; D004032:Diet; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019833:Genistein / Q000494:pharmacology; D007529:Isoflavones / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011282:Pregnenediones / Q000378:metabolism; D011375:Progesterone Reductase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013251:Steroid Isomerases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D043266:Steryl-Sulfatase; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000378:metabolism
C021425:3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase; D003374:Coumarins; D004791:Enzyme Inhibitors; D007529:Isoflavones; D009097:Multienzyme Complexes; D011282:Pregnenediones; C007768:formononetin; C004742:daidzein; D019833:Genistein; D011375:Progesterone Reductase; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D001192:Arylsulfatases; D043266:Steryl-Sulfatase; D013251:Steroid Isomerases; C004541:biochanin A