Comparative studies on binding of 3 different ligands to the N-methyl-D-aspartate recognition domain in brain...

Treatment with a low concentration of Triton X-100 almost tripled the binding of [3H]D,L-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid (CGP 39653), a novel competitive antagonist at an N-methyl-D-aspartate (NMDA)-sensitive subclass of brain excitatory amino acid receptors, in synaptic membranes of the rat brain. The binding linearly increased with increasing protein concentrations of up to 0.4 mg/ml and also increased in proportion to incubation time with a plateau within 60 min after the initiation of incubation at 2 degrees C in Triton-treated membranes. Elevation of incubation temperature from 2 degrees C to 30 degrees C resulted in a marked decrease in the binding at equilibrium by 80%, and a maximal level was obtained within 1 min after the initiation of incubation at 30 degrees C with a gradual decline of up to 10 min. Bound [3H]CGP 39653 was rapidly dissociated by the addition of excess unlabeled L-glutamic acid (Glu), and the time required to attain complete dissociation was 60 min at 2 degrees C and 1 min at 30 degrees C, respectively. Among several agonists and antagonists tested, Glu was the most potent displacer of [3H]CGP 39653 binding with progressively less potent displacement by D-2-amino-5-phosphonovaleric, (+-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic (CPP), D-2-amino-7-phosphonoheptanoic, N-methyl-D-aspartic and N-methyl-L-aspartic acids.(ABSTRACT TRUNCATED AT 250 WORDS)