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. 2021 Sep 8;120(20):4429–4441. doi: 10.1016/j.bpj.2021.09.004

Figure 5.

Figure 5

Double mutation of E216A and D1196N in CaV1.1e reduces the current density and right shifts the activation curve toward that of CaV1.1a. (A) Side and top view of structures of the PD I – VSD IV interface of CaV1.1e substitution E216A/D1196N, showing the total loss of ionic interactions, between the gating charges R1/R2 (blue) and the mutated countercharges N1196 and A216 (pink) (dashed line, remaining H-bond). (B–E) Representative example traces (B), I-V curves (C), and G-V curves (D) of calcium currents recorded from myotubes expressing wild-type GFP-CaV1.1e (red), wild-type GFP-CaV1.1a (blue), or GFP-CaV1.1e-E216A/D1196N (orange). (E) The simultaneous E216A and D1196N substitution causes a significant right shift of V1/2 of 21.6 mV (N = 5–7; p < 0.0001). No significant difference in V1/2 was observed between wild-type CaV1.1a and CaV1.1e-E216A/D1196N (p = 0.2). P-values were calculated using an ANOVA with Tukey's posthoc test, error bars indicate standard error. For further current parameters, see Table 1. To see this figure in color, go online.