THE VERTEBRATE ALCOHOL DEHYDROGENASE SYSTEM - VARIABLE CLASS II TYPE FORM ELUCIDATES SEPARATE STAGES OF ENZYMOGENESIS

摘要

A mixed-class alcohol dehydrogenase has been characterized from avian liver, Its functional properties resemble the classical class I type enzyme in livers of humans and animals by exhibiting low K-m and k(cat) values with alcohols (K-m = 0.7 mM with ethanol) and low K-i values with 4-methylpyrazole (4 mu M). These values are markedly different from corresponding parameters of class II and III enzymes, In contrast, the primary structure of this avian liver alcohol dehydrogenase reveals an overall relationship closer to class II and to some extent class III (69 and 65% residue identities, respectively) than to class I or the other classes of the human alcohol dehydrogenases (52-61%), the presence of an insertion (four positions in a segment close to position 120) as in class II but in no other class of the human enzymes, and the presence of several active site residues considered typical of the class II enzyme, Hence, the avian enzyme has mixed-class properties, being functionally similar to class I, yet structurally similar to class II, with which it also clusters in phylogenetic trees of characterized vertebrate alcohol dehydrogenases, Comparisons reveal that the class II enzyme is approximate to 25% more variable than the ''variable'' class I enzyme, which itself is more variable than the ''constant'' class III enzyme, The overall extreme variability, the activity unexpected for a class II enzyme, and the unusual chromatographic behavior may explain why the class II enzyme has previously not been found outside mammals, The properties define a consistent pattern with apparently repeated generation of novel enzyme activities after separate gene duplications. [References: 26]