RESUMO
The channel-associated protein PSD-95 functionally modulates NMDA receptor channels, interacting with the channels via PDZ domain of PSD-95. PSD-95 also interacts with guanylate kinase-associated protein (GKAP) through the guanylate kinase-like domain of PSD-95. Here we report that GKAP markedly potentiates the channel activity of the receptor-PSD-95 complex. However, GKAP had no effect on basic properties of the channels nor on PSD-95-induced changes in channel properties. Thus, GKAP affects the channel activity of the NMDA receptor via PSD-95 quantitatively, which may make signal transmission more efficient at postsynaptic sites.
Assuntos
Proteínas de Transporte/farmacologia , Proteínas do Tecido Nervoso/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Animais , Expressão Gênica , Ácido Glutâmico/farmacologia , Canais Iônicos/genética , Microinjeções , Mutação , Proteínas do Tecido Nervoso/genética , Oócitos , Técnicas de Patch-Clamp , RNA Mensageiro/genética , Receptores de N-Metil-D-Aspartato/genética , Proteínas Associadas SAP90-PSD95 , Acetato de Tetradecanoilforbol/farmacologia , Xenopus laevisRESUMO
A channel-associated protein PSD-95 has been shown to induce clustering of N-methyl D-aspartate (NMDA) receptors, interacting with the COOH terminus of the epsilon subunit of the receptors. The effects of PSD-95 on the channel activity of the epsilon2/zeta1 heteromeric NMDA receptor were examined by injection of PSD-95 cRNA into Xenopus oocytes expressing the NMDA receptors. Expression of PSD-95 decreased the sensitivity of the NMDA receptor channels to L-glutamate. Mutational studies showed that the interaction between the COOH terminus of the epsilon2 subunit of the NMDA receptor and the second PSD-95/Dlg/Z0-1 domain of PSD-95 is critical for the decrease in glutamate sensitivity. It is known that protein kinase C markedly potentiates the channel activity of the NMDA receptor expressed in oocytes. PSD-95 inhibited the protein kinase C-mediated potentiation of the channels. Thus, we demonstrated that PSD-95 functionally modulates the channel activity of the epsilon2/zeta1 NMDA receptor. PSD-95 makes signal transmission more efficient by clustering the channels at postsynaptic sites. In addition to this, our results suggest that PSD-95 plays a protective role against neuronal excitotoxicity by decreasing the glutamate sensitivity of the channels and by inhibiting the protein kinase C-mediated potentiation of the channels.
