FIGURE 5.

Binding of the PBM is essential for phosphatase activation by the VSD. Mutations that interfere with membrane binding of the PBM (R253A/R254A and R253K/R254K) abolish the depolarization-induced activation of enzymatic activity of Ci-VSPTEN16 as monitored by measuring membrane association of Akt-PH-GFP (n = 7 cells for each mutant). Experiments were done as described in the legend to Fig. 2, and data for Ci-VSPTEN16 are from Fig. 2D for comparison. The gray area indicates depolarization from −60 to +80 mV.