Displacement of DL-[3H]-2-amino-4-phosphonobutanoic acid ( [3H]APB) binding with methyl-substituted APB analogues and...

The binding of the excitatory amino acid antagonist DL-2-amino-4-phosphonobutanoic acid (DL-APB) to rat brain synaptic plasma membranes was characterized. As determined by Scatchard analysis, the binding was saturable and homogeneous with a Kd = 6.0 microM and Bmax = 380 pmol/mg of protein. The binding was dependent on the presence of Ca2+ and Cl- ions and was diminished upon freezing. The association rate constant was 6.8 X 10(-3) microM-1 min-1, and the dissociation rate constant was 2.0 X 10(-2) min-1. The L isomers of APB, glutamate, and aspartate were more potent as displacers of APB binding than the D isomers. Previously determined inhibition data obtained for APB-sensitive inputs to hippocampal granule cells are compared to the present displacement data in an attempt to identify this binding protein as the recognition site of the receptor mediating the APB-induced inhibition of synaptic transmission. With the exception of kynurenic acid, all compounds examined in both systems were more potent as displacers of APB binding than as inhibitors of synaptic transmission. This difference in potency was most pronounced for agonists at dentate granule cells. L-Glutamate, D-glutamate, and L-glutamate tetrazole were between 140- and 7500-fold more potent as displacers of DL-APB binding than as inhibitors of synaptic transmission. D-2-Amino-5-phosphonopentanoic acid and alpha-methyl-APB were between 10- and 20-fold more potent as displacers of binding.(ABSTRACT TRUNCATED AT 250 WORDS)