Transaminations catalysed by brain glutamate decarboxylase.
In addition to normal decarboxylation of glutamate to 4-aminobutyrate, glutamate decarboxylase from pig brain was shown to catalyse decarboxylation-dependent transamination of L-glutamate and direct transamination of 4-aminobutyrate with pyridoxal 5'-phosphate to yield succinic semialdehyde and pyridoxamine 5'-phosphate in a 1:1 stoichiometric ratio. Both reactions result in conversion of holoenzyme into apoenzyme. With glutamate as substrate the rates of transamination differed markedly among the three forms of the enzyme (0.008, 0.012 and 0.029% of the rate of 4-aminobutyrate production by the alpha-, beta- and gamma-forms at pH 7.2) and accounted for the differences among the forms in rates of inactivation by glutamate and 4-aminobutyrate. Rates of transamination were maximal at about pH 8 and varied in parallel with the rate constants for inactivation from pH 6.5 to 8.0. Rates of transamination of glutamate and 4-aminobutyrate were similar, suggesting that the decarboxylation step is not entirely rate-limiting in the normal mechanism. The transamination was reversible, and apoenzyme could be reconstituted to holoenzyme by reverse transamination with succinic semialdehyde and pyridoxamine 5'-phosphate. As a major route of apoenzyme formation, the transamination reaction appears to be physiologically significant and could account for the high proportion of apoenzyme in brain.
D062365:Aminobenzoates / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid; D005968:Glutamate Decarboxylase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism; D008956:Models, Chemical; D011733:Pyridoxamine / Q000031:analogs & derivatives / Q000378:metabolism; D013053:Spectrophotometry; D013552:Swine; D005680:gamma-Aminobutyric Acid / Q000031:analogs & derivatives / Q000378:metabolism
D062365:Aminobenzoates; D007527:Isoenzymes; C017856:3,5-diaminobenzoic acid; D005680:gamma-Aminobutyric Acid; D011733:Pyridoxamine; D005968:Glutamate Decarboxylase; C009338:succinic semialdehyde; C010627:pyridoxamine phosphate