Purification and molecular properties of mouse alcohol dehydrogenase isozymes
- 1 December 1983
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 137 (1-2), 139-147
- https://doi.org/10.1111/j.1432-1033.1983.tb07807.x
Abstract
Alcohol dehydrogenase isozymes from mouse liver (A2 and B2) and stomach (C2) tissues have been purified to homogeneity using triazine-dye affinity chromatography. The enzymes are dimers with similar but distinct subunit sizes, as determined by SDS/polyacrylamide gel electrophoresis: A, 43000; B, 39000, and C, 47000. Zinc analyses and 1,10-phenanthroline inhibition studies indicated that the A and C subunits each contained two atoms of zinc, with at least one being involved catalytically, whereas the B subunit probably contained a single non-catalytic zinc atom. The isozymes exhibited widely divergent kinetic characteristics. A2 exhibited a Km value for ethanol of 0.15 mM and a broad substrate specificity, with Km values decreasing dramatically with an increase in chain length; C2 also exhibited this broad specificity for alcohols but showed a Km value of 232 mM for ethanol. These isozymes also showed broad substrate specificities as aldehyde reductases. In contrast, B2 showed no detectable activity as an aldehyde reductase for the aldehydes examined, and used ethanol as substrate only at very high concentrations (greater than 0.5 M). The isozyme exhibited low Km and high Vmax values, however, with medium-chain alcohols. Immunological studies showed that A2 was immunologically distinct from the B2 and C2 isozymes. In vitro molecular hybridization studies gave no evidence for association between the alcohol dehydrogenase subunits. The results confirm genetic analyses [Holmes, Albanese, Whitehead and Duley (1981) J. Exp. Zool. 215, 151-157] which are consistent with at least three structural genes encoding alcohol dehydrogenase in the mouse and confirm the role of the major liver isozyme (A2) in ethanol metabolism.Keywords
This publication has 40 references indexed in Scilit:
- Purification and substrate specifities of three human liver alcohol dehydrogenase isoenzymesFEBS Letters, 1982
- New human liver alcohol dehydrogenase forms with unique kinetic characteristicsBiochemical and Biophysical Research Communications, 1981
- IMMUNOLOGICAL PROPERTIES OF THE HUMAN ALCOHOL DEHYDROGENASE (ADH) ISOZYMESInternational Journal of Immunogenetics, 1978
- Enzyme purification by substrate elution chromatography from procion dye—polysaccharide matricesFEBS Letters, 1976
- Double-ternary complex affinity chromatography: preparation of alcohol dehydrogenasesBiochemistry, 1976
- Alcohol dehydrogenase isozymes in adult human stomach and liver: evidence for activity of the ADH3 locusAnnals of Human Genetics, 1972
- Effect of substrate structure on the rate of the catalytic step in the liver alcohol dehydrogenase mechanismBiochemistry, 1971
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Subunit Composition of horse liver alcohol dehydrogenase isoenzymesFEBS Letters, 1969
- Antibody studies with the multiple enzymes of horse liver alcohol dehydrogenase IBiochemical and Biophysical Research Communications, 1968