Inactivation of D-serine dehydratase by alkylamines via a transimination of enzyme-linked cofactor.

The time-dependent inactivation of D-serine dehydratase by alkylamines was characterized. Evidence is presented indicating that inactivation proceeds via a transimination reaction analogous to the first step in the catalytic pathway. The product of alkylamine attack, an alkylamine pyridoxal 5'-phosphate Schiff base, readily dissociated from D-serine dehydratase to produce inactive apoenzyme. Reaction with alkylamines was shown to be a convenient way of producing apoenzyme which can be reconstituted with pyridoxal 5'-phosphate to fully active holoenzyme. Amino acids such as glycine and alanine, unlike alkylamines, did not resolve D-serine dehydratase but were competitive inhibitors of the enzyme. The inability of amino acids to resolve D-serine dehydratase from its cofactor was attributed to a failure of the amino acid-cofactor Schiff base to dissociate from the enzyme. Transimination of D-serine dehydratase with its substrate D-serine was at least 3.5 X 10(5) times faster than a nonenzymic model transimination reaction and more than 70 million times faster than the reaction of the enzyme with 2-hydroxyethylamine, indicating that the carboxyl group of the substrate is an important structural determinant for catalysis of the transimination step in the catalytic pathway. An analysis of the inhibitory effect of potassium ion on the rate and extent of inactivation of D-serine dehydratase by alkylamines indicated that K+ binding increased the affinity of the enzyme for its cofactor by at least 13-fold.