Abstact

gamma-Aminobutyric acid aminotransferase (GABA-T) is a pyridoxal 5'-phosphate (PLP)-dependent enzyme that catalyzes gamma-aminobutyric acid (GABA) degradation in the mitochondrial GABA shunt. Plant GABA-Ts correspond to EC 2.6.1.96 and differ from mammalian and bacterial GABA-Ts (EC 2.6.1.19) in substrate specificity; however, their structural basis has remained unclear. Here, we report the crystal structure of GABA aminotransferase from Arabidopsis thaliana (AtGABA-T) at 2.0 A resolution. Structural comparison using Foldseek indicates that AtGABA-T belongs to the class III aminotransferase family. Comparison with a class III omega-aminotransferase suggests that Arg423 located in the C-terminal region of AtGABA-T functions as the primary anchor for the carboxyl group of GABA, together with residues forming a tunnel-shaped substrate-access pathway. In contrast to nonplant GABA aminotransferases, which utilize a conserved N-terminal arginine for substrate recognition, AtGABA-T employs a distinct C-terminal arginine. These findings provide direct structural evidence for the classification of AtGABA-T as a class III aminotransferase and reveal a distinct mode of substrate recognition in AtGABA-T.