Bioactivation of chemopreventive selenocysteine Se-conjugates and related amino acids by amino acid oxidases novel route of metabolism of selenoamino acids.

Abstract

Several selenocysteine Se-conjugates have been shown to possess potent chemopreventive activity in animal models for chemical carcinogenesis. As a mechanism of action, beta-elimination reactions to form chemopreventive selenols, ammonia, and pyruvate has been proposed. The enzymes involved in these beta-elimination reactions, however, have been partially elucidated. Next to cysteine conjugate beta-lyases, as yet unidentified non-pyridoxal-5'-phosphate-dependent enzymes also appear to be involved in cytosolic beta-elimination reactions. In the present study, it was investigated whether amino acid oxidases contribute to the bioactivation of selenocysteine Se-conjugates. Using purified L-amino acid oxidase from Crotalus adamanteus as a model enzyme, significant beta-elimination activities were indeed observed upon incubation with Se-methylselenocysteine (K(m), 195 microM; k(cat), 48 min(-1)), Se-allylselenocysteine (K(m), 608 microM; k(cat), 34 min(-1)), Se-phenylselenocysteine (K(m), 107 microM; k(cat), 57 min(-1)) and Se-benzylselenocysteine (K(m), 59 microM; k(cat), 13 min(-1)). For all selenocysteine Se-conjugates tested, the rate of pyruvate formation was comparable to that of hydrogen peroxide, one of the products of oxidative deamination. The fact that addition of catalase did not alter pyruvate formation indicated that the beta-elimination reaction observed was not mediated by selenoxidation/syn-elimination due to the hydrogen peroxide formed via the oxidative deamination pathway. Using D-amino acid oxidase from porcine kidney and D-SeCys conjugates similar results were obtained. To delineate whether mammalian L-amino acid oxidases are also able to catalyze beta-elimination of selenocysteine Se-conjugates, rat renal cytosol was fractionated and screened for beta-elimination and oxidative deamination activities. One of the fractions isolated displayed oxidative deamination activity with several amino acids and cysteine S-conjugates. With selenocysteine Se-conjugates as substrates, however, this fraction displayed both oxidative deamination and beta-elimination activities, when incubated in the presence of aminoxyacetic acid to block contribution of pyridoxal-5'-phosphate-dependent enzymes. The potential significance of this novel bioactivation route for the chemopreventive activity of selenocysteine Se-conjugates is discussed.