title: Purification and kinetic characterization of pickerel liver alcohol dehydrogenase with dual coenzyme specificity.
abstract: A major alcohol dehydrogenase isozyme that displays dual coenzyme specificity has been purified from pickerel liver by ion-exchange, gel filtration, and affinity chromatographic procedures. The purified enzyme is chromatographically and electrophoretically homogeneous. It is dimeric and possesses common physical properties shared by other liver alcohol dehydrogenases. Phosphorus-31 nuclear magnetic resonance spectroscopy demonstrates that NADP+ binds to two coenzyme sites of the pickerel enzyme. Steady-state kinetic studies suggest that pickerel liver alcohol dehydrogenase catalyzes NAD(P)(+)-linked ethanol oxidation via a random pathway. While the NADP+ reduction involves the formation of an abortive complex at high NADP+ concentrations, the NAD+ reduction at low NAD+ concentrations follows an ordered Bi-Bi mechanism with NAD+ being the leading reactant.
authors.affiliation: Department of Chemistry, Carleton University, Ottawa, ON, Canada.
authors.forename: L S
authors.initials: LS
authors.lastname: al-Kassim
authors.forename: C S
authors.initials: CS
authors.lastname: Tsai
mesh_terms: D000426:Alcohol Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D017859:Esocidae; D007526:Isoelectric Point; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D009243:NAD / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*
chemical_list: D007527:Isoenzymes; D046911:Macromolecular Substances; D009243:NAD; D009249:NADP; D000426:Alcohol Dehydrogenase