Biochemical identification of a putative glutamate receptor in housefly thoracic membranes

Specific stereoselective binding of 3H]L-glutamate was detected to membranes prepared from housefly thorax to which were added several antiproteases. A single high affinity binding site was detected (KD 0.5 +/- 0.04 microM), but total binding varied from preparation to preparation (5-60 pmoles/mg protein). Specific binding was inhibited by preincubation of the membranes with trypsin, chymotrypsin or protease, or by exposure to 70 degrees C for 5 min. It was also inhibited by several compounds, the most potent being L-glutamate and L-aspartate, followed by L-glutamate diethylester, then D-glutamate, N-methyl-D-aspartate and ibotenate. Quisqualate had little effect, while kainate, proctolin and D-aspartate had none. d-Tubocurarine stimulated 3H]L-glutamate binding. The data suggest that 3H]L-glutamate is binding to an L-glutamate receptor in housefly thoracic muscle membranes.