Mutagenesis experiments show the importance of interacting residues from simulations. Experiments were done on human KCNQ1 channel with the mutations Y281F, Y278F, and K285Q; the Xenopus numberings Y271F, Y268F, and K275Q were used here for ease of comparison with the simulations. (A) ΔV0.5 dose response curve of LIN-Gly on WT and point mutation KCNQ1 channels (WT n = 10, Y271F n = 3, Y268F n = 4, K275Q n = 5). (B) ΔV0.5 dose response curve of LIN-Tyr on WT and point mutation KCNQ1 channels (WT n = 6, Y271F n = 4, Y268F n = 4, K275Q n = 3). (C) Comparison of ΔV0.5 effect on WT KCNQ1 channels versus point mutations at 7 µM of LIN-Gly (WT: −25.28 ± 1.69 mV; n = 10, Y271F: −25.49 ± 2.53 mV; n = 3, Y268F: −13.91 ± 1.7 mV; n = 4, K275Q: −7.66 ± 3.04 mV; n = 5). ANOVA with Tukey’s multiple comparison of WT to each point mutation gave ns for Y271F, P = 0.067 for Y268F and P = 0.0006 for K275Q. ***, P < 0.001. (D) Comparison of ΔV0.5 effect on WT KCNQ1 channels versus point mutations at 7 µM of LIN-Tyr (WT: −43.57 ± 1.03 mV; n = 3, Y271F: −52.28 ± 2.98 mV; n = 4, Y268F: −48.23 ± 1.15 mV; n = 4, K275Q: −39.47 ± 14.8 mV; n = 3). ANOVA with Tukey’s multiple comparison of WT to each point mutation gave ns for all comparisons. Data shown as mean ± SEM.