Figure 1.

S343D and S344D mutations in the CaMBD of CCaMK did not alter the interactions between calmodulin (CaM) and the peptides. Mobility shift assay using oligo-peptides covering the CaMBD (30 amino acids in length): equal mole of peptide and CaM (1ug/per reaction) was incubated in reaction buffer (25mM Tris-HCl, 150mM NaCl and 1mM CaCl₂, pH7.5) for 15 minutes at room temperature and run in non-denaturing polyacrylamide gel in the presence of 1mM CaCl₂. The gel was then stained with Coomassie solution until bands were observed. Small peptides including a phosphomimic mutation at Ser-343 and Ser-344 interacted with CaM.