FIGURE 3.

Binding site for CaMKII on Ca_V2.1 channels. A, binding of CaMKII was detected by GST pulldown assay. CaMKII (20 nm) was used as prey in the presence/absence of Ca²⁺/CaM (5 μm) and ATP (1 mm) as indicated. GST alone or GST-tagged Ca_V2.1(1848–1964) was used as bait as indicated: WT, ¹⁹⁰³TVGKIY¹⁹⁰⁸; mutant, ¹⁹⁰³EEDAAA¹⁹⁰⁸. a, the blot was probed with anti-phospho-CaMKII(Thr-286). b, the same blot after stripping and re-probing using anti-GST to show equal loading of Ca_V2.1(1848–1964). B, quantitation of relative CaMKII autophosphorylation using anti-phospho CaMKII(Thr-286) under the indicated conditions is shown (mean ± S.E.; **, p < 0.01 by Student's t test; n = 6).