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. Author manuscript; available in PMC: 2010 Feb 10.
Published in final edited form as: Cell. 2010 Jan 22;140(2):222. doi: 10.1016/j.cell.2009.12.055

Figure 2. DAPK1 Directly Binds to NMDA Receptor NR2B Subunit.

Figure 2

(A) Representation illustrates a series of GST fusion proteins with the NR2B C-terminal mutants. (B) Affinity binding assay shows that flag-tagged catalytic domain of DAPK1 binds with GST-NR2B839–1482 (~100 kDa) and GST-NR2B1292–1304 (~30 kDa). The blots show anti-Flag (top) and anti-GST (bottom).

(C) NR2BCT suppresses DAPK1 association with NMDA receptor NR2B subunit. Membrane extracts (500 μg proteins) from the forebrain of mice 2 hr after MCAO were incubated with GST or the GST-NR2BCT839–1482 in the presence of 50 μM NR2BCT or sNR2BCT and probed with anti-DAPK1.

(D) NR2BCT blocks NMDA receptor association with DAPK1 but not with PSD-95. Membrane extracts were precipitated with nonspecific IgG (IgG) or anti-NR2B in the presence of 50 μM NR2BCT or sNR2BCT and probed with anti-DAPK1, or anti-PSD-95, or anti-CaMK-IIα. Input: 50 μg of protein was loaded in each lane. In (B)–(D), similar results were observed in each of the four experiments.