Phosphorylation-dependent interaction of the N-methyl-D-aspartate receptor ε2 subunit with phosphatidylinositol 3-kinase

Background: The NMDA receptors (NMDARs) are ion channels through which Ca2+ influx triggers various intracellular responses. Tyrosine phosphorylation of NMDARs regulates NMDA channel activities, which may be important in neuronal plasticity. The biological significance of the tyrosine phosphorylation events, however, differs among NMDAR subunits: tyrosine phosphorylation of NMDAR epsilon 1 increases NMDA channel activities, but that of NMDAR epsilon 2 does not. Since signal transductions from various cell surface receptors are mediated by protein-protein interaction through phosphotyrosine and the Src homology 2 (SH2) domain, we examined the possibility that phosphotyrosines in NMDAR epsilon 2 contribute to the intracellular signalling events. Results: We first show that Fyn is deeply involved in the phosphorylation of NMDAR epsilon 2 and second that a phosphotyrosine in NMDAR epsilon 2 interacts with the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3-kinase). Both the level of tyrosine phosphorylation on NMDAR epsilon 2 and the amounts of the p85 subunit (p85) bound to NMDAR epsilon 2 are decreased in Fyn-deficient mice. Moreover, we show that ischaemia stimulates the binding of p85 to phosphorylated NMDAR epsilon 2, suggesting a physiological role of the phosphotyrosine/SH2-based interaction between NMDAR epsilon 2 and p85 in the brain. Conclusions: The tyrosine phosphorylation event on NMDARs is important in not only the regulation of its channel activity but also intracellular signalling mediated through the interaction of the NMDAR with SH2 domain-containing molecules.