title: Characterization of the half and overall reactions catalyzed by L-lysine:2-oxoglutarate 6-aminotransferase.
abstract: Significant differences were found in the reaction rate, and the substrate and reaction specificities between the half reactions and the overall reactions catalyzed by L-lysine: 2-oxoglutarate 6-aminotransferase. The half reactions between an amino donor and the enzyme-bound pyridoxal 5'-phosphate, and also between an amino acceptor and the bound pyridoxamine 5'-phosphate followed first order reaction kinetics. The extrapolated first order rate constants and dissociation constants of the substrates were determined for the half reactions: lysine, 0.87 min-1 and 5.5 mM; glutamate, 1.1 min-1 and 10.5 mM; alanine, 0.66 min-1 and 6.6 mM; 6-aminohexanoate, 0.43 min-1 and 13.3 mM; and 2-oxoglutarate, 0.33 min-1 and 2.5 mM. As compared with the values reported for the overall reactions [Soda, K., Misono, H., & Yamamoto, T. (1968) Biochemistry 7, 4102-4109], the reactivity of the inherent substrates was lower by over 4 orders in the half reaction than that in the overall reaction, and the reactivity of alanine with the bound pyridoxal 5'-phosphate was reduced to 10% of that in the overall reaction. The substrate specificity in the half reaction was much lower than that in the overall reaction, which was re-examined in a reaction system containing the same concentration of the enzyme as that for the half reactions. Lysine 6-aminotransferase catalyzes the transfer of only the terminal amino group of lysine to 2-oxoglutarate in the overall reaction. However, in the half reaction, the 2-amino group as well as the terminal one was transferred to the bound pyridoxal 5'-phosphate. The ratio of reactivity of the 2-amino group to that of the 6-amino group was considerably influenced by the pH of the reaction mixture.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Laboratory of Microbial Biochemistry, Kyoto University.
authors.forename: T
authors.initials: T
authors.lastname: Yagi
authors.forename: H
authors.initials: H
authors.lastname: Misono
authors.forename: K
authors.initials: K
authors.lastname: Tanizawa
authors.forename: T
authors.initials: T
authors.lastname: Yoshimura
authors.forename: K
authors.initials: K
authors.lastname: Soda
mesh_terms: D001665:Binding Sites; D005417:Flavobacterium / Q000201:enzymology; D007700:Kinetics; D051306:L-Lysine 6-Transaminase; D008239:Lysine; D011732:Pyridoxal Phosphate; D013379:Substrate Specificity; D000637:Transaminases / Q000378:metabolism*
chemical_list: D011732:Pyridoxal Phosphate; D000637:Transaminases; D051306:L-Lysine 6-Transaminase; D008239:Lysine