Purification and characterization of mouse alcohol dehydrogenase from two inbred strains that differ in total liver enzyme activity.

Alcohol dehydrogenase activity in mouse liver homogenate-supernatants is 1.7 times greater in the C57BL/10 strain than in the BALB/c strain, regardless of whether activity is expressed in units per gram liver, total liver, or milligram DNA. The Km values for ethanol and NAD+, approximately 0.4 and 0.03 mM, respectively, of enzyme purified from both strains are similar. Moreover, the Ki for NADH, 1 microM, the pH optimum for ethanol oxidation, 10.5, and the Vmax for ethanol oxidation, 160 min-1, for ADH from the C57BL/10 and BALB/c strains are similar. Therefore, the difference in ADH activity in the two strains cannot be due to differences in the catalytic properties of the enzyme. The electrophoretic and isoelectric focusing patterns and two-dimensional tryptic peptide maps of the purified enzyme from both strains are identical. Thus the amino acid sequences of enzyme from C57BL/10 and BALB/c mice must also be identical or very similar. The difference in ADH activity in the two strains is most likely the result of genetic differences in the content of ADH protein in liver.