N-Acetylaspartylglutamate catabolism is achieved by an enzyme on the cell surface of neurons and glia

Abstract

N-Acetylaspartylglutamate (NAAG) is a nervous system-specific, acidic dipeptide which is released from neurons in a manner consistent with a function in synaptic neurotransmission. The hydrolysis of NAAG to produce glutamate and N-acetylaspartate was analyzed in cell cultures prepared from murine brain cells. Peptidase activity against NAAG was found in cultures which contained both neurons and glia, as well as in cultures of glia alone. Several lines of evidence were obtained in support of the hypothesis that this peptidase activity is predominantly bound to the extracellular face of the plasma membranes of these cells. Glutamate released from NAAG accumulated in the extracellular medium. Extracellular application of peptidase inhibitors effectively reduced NAAG hydrolysis. Peptidase activity was not secreted into the cell culture medium by intact cells, and lysed cells did not release detectable peptidase activity beyond that obtained with intact cells. Replacement of extracellular sodium with choline inhibited peptide uptake while stimulating apparent extracellular NAAG hydrolysis by intact cells in culture. Finally, the steady rise in extracellular glutamate as a consequence of NAAG hydrolysis by these brain cells, including glia, supports the conclusion that glutamate uptake is not tightly coupled to peptidase activity and thus that NAAG serves as a significant source of glutamate in the synaptic space following depolarization-induced peptide release.