Starch gel electrophoresis of homogenates from human stomach mucosa resolves three alcohol dehydrogenase (ADH) forms: the anodic χ-ADH (class III), the cathodic γ-ADH (class I), and a new form of slow cathodic mobility that has not been previously characterized. In this work, we describe the purification in three chromatographic steps and the physical and kinetic characterization of this new human alcohol dehydrogenase, which we have named σ-ADH. The enzyme exhibits the general physicochemical features (Mr, zinc content, subunit Mr, cofactor preference) of all mammalian alcohol dehydrogenases. The kinetic studies show a high Km value (41 HIM) and a high kcat value (280 min-1) for ethanol at pH 7.5. The Km decreases as the alcohol increases its chain length. The aldehydes are better substrates than the corresponding alcohols, with m-nitrobenzaldehyde being the best substrate examined. σ-ADH is strongly inhibited by 4-methylpyrazole, but with a Ki (10 μM) still higher than that for a class I isoenzyme. These properties suggest that σ-ADH is a class II isoenzyme, different from π-ADH and similar to that previously described by us in rat stomach. At the high ethanol concentrations in stomach after drinking, σ-ADH is probably the ADH form with the largest contribution to human gastric ethanol metabolism.
|Journal||Journal of Biological Chemistry|
|Publication status||Published - 15 Jan 1991|