3 beta-Hydroxy-5 beta-steroid dehydrogenase activity of human liver alcohol dehydrogenase is specific to gamma-subunits.
Human liver alcohol dehydrogenase [alcohol:NAD+ oxidoreductase, EC 1.1.1.1 (ADH)] catalyzes the stereospecific oxidation of different 3 beta-hydroxy-5 beta-steroids with ranges of Km from 46 to 320 microM and values of kcat from 7.0 to 72 min-1, pH 8.5. Only the class I isozymes containing gamma-subunits, gamma 1 gamma 1, alpha gamma 1, beta 1 gamma 1, gamma 2 gamma 2, and beta 1 gamma 2, catalyze oxidation of these steroids with kcat/Km ratios 4-10-fold greater than those for ethanol. In marked contrast, class I alpha alpha, alpha beta 1, and beta 1 beta 1, class II, and class III isozymes do not oxidize 3 beta-hydroxy-5 beta-steroids though they readily oxidize ethanol. 1,10-Phenanthroline and 4-methylpyrazole competitively inhibit both alcohol dehydrogenase catalyzed ethanol and 3 beta-hydroxy-5 beta-steroid oxidation demonstrating that the catalysis of both types of substrates occurs at the same active site. The gamma-subunit-catalyzed oxidation of 3 beta-hydroxy-5 beta-steroids is the most specific catalytic function described thus far for any human liver alcohol dehydrogenase isozyme: there is no other isozyme that catalyzes this reaction. Testosterone, an allosteric inhibitor of ethanol oxidation specific for gamma-subunit-containing human liver ADH isozymes [Mårdh, G., Falchuk, K. H., Auld, D. S., & Vallee, B. L. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 2836-2840], also noncompetitively inhibits gamma-subunit-catalyzed sterol oxidation.(ABSTRACT TRUNCATED AT 250 WORDS)
McEvily
A J
AJ
Center for Biochemical and Biophysical Sciences and Medicine, Harvard Medical School, Boston, Massachusetts 02115.
Holmquist
B
B
Auld
D S
DS
Vallee
B L
BL
D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008099:Liver / Q000201:enzymology*
D007527:Isoenzymes; D015096:3-Hydroxysteroid Dehydrogenases; D000426:Alcohol Dehydrogenase