Kinetic mechanisms in the reduction of aldehydes and ketones catalyzed by rabbit liver aldehyde reductases and hydroxysteroid dehydrogenases.

The kinetic properties of the NADPH-dependent reduction of aromatic aldehydes and ketones catalyzed by low- and high-molecular-weight aldehyde reductases [alcohol : NADP oxidoreductase, EC 1.1.1.2] and 3 alpha- and 3 beta-hydroxysteroid dehydrogenases [EC 1.1.1.50 and 1.1.1.51] of rabbit liver were compared. Initial velocity measurements with pyridine-4-aldehyde, 4-benzoylpyridine and androstadione as substrates and inhibition studies with their products indicated that all the enzymes followed an ordered Bi Bi reaction mechanism with coenzyme binding first and leaving last. However, phenylpyruvic acid inhibited 3 alpha-hydroxysteroid dehydrogenase and low-molecular-weight aldehyde reductase noncompetitively with respect to either NADPH or substrate, whereas it inhibited 3 beta-hydroxysteroid dehydrogenase and high-molecular-weight aldehyde reductase uncompetitively. Cibacron blue F3GA dye was a dead-end inhibitor of the enzymes, being competitive with respect to NADPH and noncompetitive with respect to the other substrate, but the Ki value of 3 alpha-hydroxysteroid dehydrogenase for this dye was much higher than those of the other enzymes.