Biosynthesis of 4-aminobutyrate aminotransferase.

Mitochondrial 4-aminobutyrate aminotransferase was synthesized in a cell-free reticulocyte lysate using polysomal RNA isolated from pig brain. Its primary translation product has a higher molecular mass than the mature enzyme. The difference in relative molecular mass is approximately 2000 as revealed by SDS/polyacrylamide gel electrophoresis. The precursor of 4-aminobutyrate aminotransferase recognizes polyvalent antibodies raised against the mature enzyme. The precursor of 4-aminobutyrate aminotransferase binds pyridoxal-5-P and displays catalytic activity. Enzymatic activity was detected using a sensitive fluorimetric method, which is based on the formation of condensation products between succinic semialdehyde and cyclohexane-1,3-dione. It is concluded that removal of an extra peptide from the precursor is not an obligatory first step in the production of biological active species.