Kinetics of the Sodium‐dependent Transport of Gamma‐aminobutyric Acid by Synaptosomes

The kinetics of transport of gamma‐aminobutyric acid [2,3‐3H] by synaptosomes from rat brain was studied by means of a rapid filtration technique. The rate of uptake was proportional to the protein concentration over the range 0.05—0.2 mg of synaptosomal protein per ml. Although apparent allosteric kinetics were observed with sodium, transport followed simple saturation kinetics with respect to GABA and no heterotropic, cooperative effects of GABA on sodium on kinetics were observed. A minimum of three interacting sodium sites is suggested the basis of Hill plots of the sodium data. Both the apparent Km and Vmax for GABA were functions of the sodium ion concentration but the effect of sodium was considerably greater on Vmax than on the apparent Km The Vmax for GABA was 1.1 ± 0.5 nmol.min−1 mg−1 of protein at 95 mm sodium and decreased to 12 per Cent of this value at 19 mm sodium. The apparent Km for GABA increased from 4.0 ± 1.0 μm at 95 mm sodium to 8.4 ± 2.0 μm at 19 mm sodium. Potassium was a noncompetitive inhibitor with respect to GABA and did not affect the apparent cooperativity observed with sodium. These findings are discussed in terms of models of GABA transport.