FIGURE 1.

The phospho-mimetic GluN2B S1303D mutation impairs CaMKII binding to GluN2B. A, binding of purified CaMKIIα to immobilized GST-N2B-c (a GST-fusion with the cytoplasmic C terminus of GluN2B) was induced by Ca²⁺/CaM (Ca²⁺) or by pre-autophosphorylation at T286 (P), both for N2B-c wild type and S1303A, but to a much lesser extent for the phospho-mimetic S1303D mutant. Bound CaMKII (and immobilized GST-N2B-c) was eluted and detected by Western blot analysis. B, quantification of binding experiments as shown in panel A (n = 3). CaMKII bound similarly to N2B-c wild type and S1303A (n.s.) but significantly less to S1303D (**, p < 0.01; *, p < 0.05; in Neuman-Keuls multiple comparison test, after one-way ANOVA), both for CaM-induced and phospho-T286-induced binding (each normalized for binding to N2B wild type; IDV of phospho-T286-induced binding was 1.3–4.0-fold higher in the individual experiment series). Error bars show S.E.