Purification and partial amino acid sequence of a high-activity human stomach alcohol dehydrogenase

Journal Article

To understand the relative importance of alcohol dehydrogenase (ADH) isoenzymes in gastric ethanol metabolism, a stomach-specific ADH (σ-ADH) was purified to homogeneity from human transplant donor and surgical tissues, and its activity for ethanol oxidation was examined. The enzyme from these tissues had a specific activity at pH of ~70 units/mg, about 10 times that reported by Moreno and Pares (J. Biol. Chem. 266:1128-1133, 1991). The enzyme exhibited a high K(m) for ethanol at pH 7.5 and 10 (29 and 5.2 mM, respectively). This high-activity σ-ADH isoenzyme migrated on starch and isoelectric focusing gels to a position slightly anodic to the liver ππ isoenzyme. It was subjected to digestion by endoproteinases, and ~40% of the protein was sequenced. The σ-ADH exhibited 75%, 68%, and 62% sequence identity to the human class I (β1), II (π), and III (χ) isoenzymes, respectively, and 61% identity to the deduced ADH6 amino acid sequence. Phylogenetic analysis indicated that precursors to this high-activity σ-ADH and the class I isoenzymes diverged more recently than precursors to the class II and III isoenzymes, after reptilian and avian divergence. The high- activity σ-ADH isoenzyme therefore represents a distinct class of ADH (class IV), more closely related in evolution to the class I isoenzymes than to the other known human isoenzymes.

Cited Authors

  • Stone, CL; Thomasson, HR; Bosron, WF; Li, TK

Published Date

  • 1993

Published In

  • Alcoholism: Clinical and Experimental Research

Volume / Issue

  • 17 / 4

Start / End Page

  • 911 - 918