The effects of pH on the rates of isotope exchange catalyzed by alanine aminotransferase.

Alanine aminotransferase catalyzes exchange of the β-hydrogens of alanine with the solvent at a rate commensurate with the rate of exchange of the α-hydrogen. These methyl protons are lost sequentially and intermediates having protons on the α-carbon but deuterium on the β-carbon were detected by nuclear magnetic resonance. The overall rates of exchange of both α-hydrogen and β-hydrogen were less than the rate of transamination and did not vary from pH 6–8. The α-hydrogen of glutamate, on the other hand, was found to exchange at a greater rate than the overall transamination rate with ketoglutarate. However the β-hydrogens of glutamate are not removed during the enzymic reaction. It is concluded that a basic group on the enzyme removes the proton from the α-carbon of alanine at a rate at least as great as the rate of transamination. Because the proton is held on the enzyme, it appears to exchange more slowly in alanine. Labilization of the α-hydrogen of amino acids does not appear to be the ratelimiting reaction of alanine aminotransferase, but occurs at a rate comparable to that of the overall reaction.