Salek, Asma B.Edler, Michael C.McBride, Jonathon P.Baucum, Anthony J.2021-08-092021-08-092019-08-02Salek, A. B., Edler, M. C., McBride, J. P., & Baucum, A. J. (2019). Spinophilin regulates phosphorylation and interactions of the GluN2B subunit of the N-methyl-d-aspartate receptor. Journal of Neurochemistry, 151(2), 185–203. https://doi.org/10.1111/jnc.148311471-4159https://hdl.handle.net/1805/26358N-methyl-D-Aspartate receptors (NMDARs) are abundant postsynaptic proteins that are critical for normal synaptic communication. NMDAR channel function is regulated by multiple properties, including phosphorylation. Inhibition of protein phosphatase 1 in hippocampal neurons increases NMDAR activity, an effect abrogated by loss of spinophilin, the major protein phosphatase 1 (PP1)-targeting protein in the postsynaptic density (PSD). However, how spinophilin regulates PP1-dependent NMDAR function is unclear. We hypothesize that spinophilin regulates PP1 binding to the NMDAR to alter NMDAR phosphorylation. Our data demonstrate that spinophilin interacts with the GluN2B subunit of the NMDAR. In HEK293 cells, activation and/or overexpression of protein kinase A increased the association between spinophilin and the GluN2B subunit of the NMDAR. Functionally, we found that spinophilin overexpression decreased PP1 binding to the GluN2B subunit of the NMDAR and attenuated the PP1-dependent dephosphorylation of GluN2B at Ser-1284. Moreover, in P28 hippocampal lysates isolated from spinophilin KO compared to WT mice, there was increased binding of GluN2B to PP1, decreased phosphorylation of GluN2B at Ser-1284, and altered GluN2B protein interactions with PSD-enriched proteins. Together, our data demonstrate that spinophilin decreases PP1 binding to GluN2B and concomitantly enhances the phosphorylation of GluN2B at Ser-1284. The putative consequences of these spinophilin-dependent alterations in GluN2B phosphorylation and interactions on synaptic GluN2B localization and function are discussed.en-USProtein PhosphorylationProtein Phosphatase 1PostsynapticSignalingProtein-protein interactionsProteomicsArticle