Figure 9.

Aspartate substitution at Thr-627 in combination with aspartate substitutions at the major sites did not affect calmodulin binding. Calmodulin binding curves for WT (open circles, four data sets), triple mutant (open diamonds, three data sets), and triple mutant after 20 h PAK treatment (solid diamonds, two data sets) are shown with their 95% confidence limits (dotted lines). The binding curve for the triple mutant (K_d = 1.5 μM) was similar to that of CaD22DD (short dashed line, K_d=1.6 μM). PAK phosphorylation of the triple mutant reduced the affinity to 4.5 μM, which is similar to that observed for CaD22WT-PAK (vertical bar line). Conditions are the same as in Fig. 3.