Persistence of binding of autophosphorylated CaMKII to CaV2.1(1848–1964).
A, binding of CaMKII was detected by GST pulldown assay. CaV2.1(1848–1964) was incubated with CaMKII (20 nm) and Mg2+(5 mm) in the presence/absence of Ca/CaM (5 μm) and ATP (1 mm) as indicated. Samples were washed in the absence or presence of 5 mm EGTA as indicated. a, the blot was probed with anti-CaMKII antibody. b, the same blot after stripping and re-probing using anti-phospho-CaMKII(Thr-286). c, the same blot after stripping and reprobing using anti-CaM. d, the same blot after stripping and re-probing using anti-GST to show equal loading of CaV2.1(1848–1964). B, quantitation of relative CaMKII binding using anti-CaMKII under the indicated conditions (mean ± S.E.; **, p < 0.01; ***, p < 0.001 by Student's t test; n = 4).