Substrate specificity of human class I alcohol dehydrogenase homo- and heterodimers containing the beta 2 (Oriental) subunit.
In order to gain a better understanding of the metabolism of ethanol in Orientals, the kinetic properties of human alcohol dehydrogenase (ADH) isozymes containing the beta 2 (Oriental) subunit, i.e., alpha beta 2, beta 2 gamma 1, beta 2 beta 2, beta 2 gamma 2, as well as gamma 1 gamma 1, were examined by using primary and secondary alcohol substrates of various chain lengths and compared with those of the corresponding beta 1 (Caucasian) subunit containing isozymes already on record [Wagner, F. W., Burger, A. R., & Vallee, B. L. (1983) Biochemistry 22, 1857-1863]. With primary alcohols, these isozymes follow typical Michaelis-Menten kinetics with a preference for long-chain alcohols, as indicated by Km and kcat/Km values. The kcat values obtained with primary alcohols, except methanol, do not vary greatly, i.e., less than 3-fold, whereas the corresponding Km values span a 3600-fold range, i.e., from 26 microM to 94 mM, indicating that the specificity of these isozymes manifests principally in substrate binding. As a consequence, ethanol--which might be thought to be the principal in vivo substrate for ADH--is oxidized rather poorly, i.e., from 50- to 90-fold less effectively than octanol. Secondary alcohol oxidation by the homodimers beta 2 beta 2 and gamma 1 gamma 1 also follows normal Michaelis-Menten kinetics. Again, values of Km and kcat/Km reveal that both isozymes prefer long carbon chains. For all secondary alcohols studied, the Km and kcat values for beta 2 beta 2 are much higher than those for gamma 1 gamma 1, i.e., 25- to 360-fold and 6- to 16-fold, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Fong
W P
WP
Department of Biochemistry, Chinese University of Hong Kong, Shatin, N.T.
Keung
W M
WM
D000426:Alcohol Dehydrogenase / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D011485:Protein Binding; D013379:Substrate Specificity
D007527:Isoenzymes; D046911:Macromolecular Substances; D000426:Alcohol Dehydrogenase