In vitro studies of human liver alcohol dehydrogenase variants using a variety of substrates.
Alcohol dehydrogenase (ADH) is genetically polymorphic, and large differences in allele frequencies exist between the major human races. Genetic variants at the ADH2 gene locus include the beta 2-ADH ("atypical" ADH) present in 85% of Orientals and the beta 1-ADH ("normal" ADH) present in 85 to 95% of whites. Although the presence of one or the other of these ADH variants does not significantly affect the rate of ethanol oxidation in the living subject, it may affect that of other substrates. The overall objective of this work was to screen in vitro for ADH substrates which might be differentially metabolized by these ADH2variants in living subjects. In an in vitro screening method using autopsy livers at pH 8.5, the formation or disappearance of NADH at 340 nm was measured before and after exposure to 4-methylpyrazole, an ADH-specific competitive inhibitor. The screening test revealed three new substrates and suggested that alcohol substrates fall into two groups. The majority of substrates belonged to a group which was oxidized at a significantly lower rate by the beta 2-ADH as compared to the beta 1-ADH, but this was not the case for a small group which included ethanol. Subsequent kinetic studies of selected alcohols tended to indicate a uniqueness of ethanol kinetics in that both KM and Vmax favored its oxidation by beta 2-ADH rather than by the beta 1 variant. None of the other seven tested alcohols showed a similar differential. Also, reduction of aldehydes and ketones tended to be moderately slower by beta 2- than by beta 1-ADH.(ABSTRACT TRUNCATED AT 250 WORDS)
Kassam
J P
JP
Department of Pharmacology, University of Toronto, Ontario, Canada.
Tang
B K
BK
Kadar
D
D
Kalow
W
W
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000426:Alcohol Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D000447:Aldehydes; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D009243:NAD / Q000378:metabolism; D011110:Polymorphism, Genetic; D013379:Substrate Specificity
D000447:Aldehydes; D007527:Isoenzymes; D009243:NAD; D000426:Alcohol Dehydrogenase