Evidence for compartmentalization of glutamate in rat brain synaptosomes using the glutamate sensitivity of phosphate-activated glutaminase as a functional test.

The glutamate content of rat brain synaptosomes was measured by high performance liquid chromatography to be 39 micromol/g protein. If uncompartmentalized this glutamate (4 mM) would inhibit phosphate-activated glutaminase considerably. Since the action of any endogenous effector on the enzyme is assumed to be negligible following disruption of the synaptosomes, due to dilution with the incubation medium, the inhibition by glumate and activation by phosphate were compared in intact and disrupted synaptosomes. The inhibition by endogenous glutamate in intact synaptosomes was found to correspond to less than that of 0.5 mM of added glutamate to disrupted synaptosomes, indicating that the major fraction of synaptosomal glutamate is compartmentalized.