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W55 mutations at single agonist sites. (A) The γW55A mutation hardly affects unliganded gating (Top) but substantially reduces the liganded gating (Bottom), because of a loss in favorable ΔGB1ACh (βL262S + δL265S + δP123R background; Vm = +70 mV; open is up). (B) The change in ΔGB1ACh consequent to W55A and F mutations at αγ, αδ, and αε sites (positive is a loss of favorable free energy, which was greatest at αγ; SI Appendix, Table S3).
