title: The effects of assay temperature on the complex kinetics of acetaldehyde oxidation by aldehyde dehydrogenase from human erythrocytes.
abstract: Several studies have shown preparations of the cytosolic aldehyde dehydrogenase (EC 1.2.1.3) from sheep and human liver and from human erythrocytes to exhibit complex kinetic behaviour in which the dependence of the initial velocity on the concentration of acetaldehyde gives rise to downwardly curving double-reciprocal plots. This behaviour has often been analysed in terms of a sharp discontinuity in the double-reciprocal plots and its possible implications for the oxidation of acetaldehyde and other pharmacologically important aldehydes has been a subject of speculation. In the present work, it is shown that the purified, apparently homogeneous, enzyme from human erythrocytes exhibits such complex kinetic behaviour when initial rates are determined at 25 degrees, although the double-reciprocal plots describe a smooth curve with no sharp discontinuity. However, when the assays were performed at 37 degrees there was no significant deviation from Michaelis-Menten kinetics over a wide range of acetaldehyde concentrations (0.2-30 mM). At higher concentrations of acetaldehyde inhibition occurred which was competitive with respect to NAD+. These results, which indicate that the complex kinetic behaviour of aldehyde dehydrogenase is not important at physiological temperature, are interpreted in terms of the mechanisms that have been advanced to explain the phenomena.
authors.affiliation: Department of Biochemistry, Trinity College, Dublin, Ireland.
authors.forename: G T
authors.initials: GT
authors.lastname: Henehan
authors.forename: K F
authors.initials: KF
authors.lastname: Tipton
mesh_terms: D000079:Acetaldehyde / Q000378:metabolism* / Q000494:pharmacology; D000444:Aldehyde Dehydrogenase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D010084:Oxidation-Reduction; D013696:Temperature
chemical_list: D000444:Aldehyde Dehydrogenase; D000079:Acetaldehyde