Figure 1

Characterization of αCaMKII fusion proteins. (A) Schematic representation of αCaMKII fusion proteins. (B) Kinase activities were measured using [γ-³²P]-ATP incorporated into autocamtide-2 using extracts of COS-1 cells expressing WT-αCaMKII or αCaMKII fusion proteins in the absence or presence of Ca²⁺/CaM and EGTA (n = 3). (C) Auto-phosphorylation at T286 of αCaMKII fusion proteins. COS-1 cells expressing WT-αCaMKII or αCaMKII fusion proteins were incubated with ionomycin (0.5 μM) or DMSO and then Western blotting performed using phospho-T286 αCaMKII or αCaMKII specific antibody, respectively. The graph shows relative ratio of phospho-αCaMKII/αCaMKII level (n = 3). (D) Complex forming ability of αCaMKII fusion proteins. COS-1 cells expressing WT-αCaMKII or αCaMKII fusion proteins were examined by gel filtration. Left graph represents absorbance curves at 280 nm. Right panel shows Western blot analyses assaying the amount of αCaMKII fusion proteins in each fraction (fraction No. 32–44).