Coupled enzymatic assay for estimation of branched-chain L-amino acid aminotransferase activity with 2-oxo acid substrates

A convenient continuous spectrophotometric assay for estimation of branched-chain L-amino acid aminotransferase activity was established: Branched-chain a-ore acid-dependent transamination of L-glutamate was coupled - via 2-oxoglutarate - to L-aspartate aminotransferase plus L-malate dehydrogenase or to L-alanine aminotransferase plus L-lactate dehydrogenase as indicator systems. The rate of transamination can be monitored specifically by measuring the decrease in NADH absorbance at 334 nm over time. The method was applied, e.g., for evaluation of some kinetic properties of the rat heart (iso)enzyme. (C) 1996 Academic Press, Inc.