Rabbit liver class III alcohol dehydrogenase: a cathodic isoform with formaldehyde dehydrogenase activity

Electrophoresis of rabbit liver homogenate on starch gel followed by activity staining revealed multiple forms of alcohol dehydrogenase (ADH) which, based on their electrophoretic mobilities, had been tentatively labeled as class "I," class "II," and class "III" ADHs. The class II enzyme has now been purified to homogeneity by ion exchange and affinity chromatography and, except for an isoelectric point of 7.7, closely resembles human class III ADH. It is a homodimer of molecular weight near 80,000 with a similar amino acid composition and comparable kinetic parameters for the oxidation of primary alcohols. Like the rat, human, and Escherichia coli class III ADHs, the rabbit enzyme is a glutathione-dependent formaldehyde dehydrogenase, and catalyzes the oxidation of S-hydroxymethylglutathione and the hemithiolacetal of 8-thiooctanoic acid. Ethanol up to 3 M does not saturate the enzyme, whereas longer chain primary alcohols exhibit Michaelis-Menten kinetics.