Gating Currents in the Hv1 Proton Channel
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The Hv1 proton channel shares striking structural homology with fourth transmembrane helical segment-type voltage-sensor (VS) domains but manifests distinctive functional properties, including a proton-selective “aqueous” conductance and allosteric control of voltage-dependent gating by changes in the transmembrane pH gradient. The mechanisms responsible for Hv1's functional properties remain poorly understood, in part because methods for measuring gating currents that directly report VS activation have not yet been described. Here, we describe an approach that allows robust and reproducible measurement of gating-associated charge movements in Hv1. Gating currents reveal that VS activation and proton-selective aqueous conductance opening are thermodynamically distinct steps in the Hv1 activation pathway and show that pH changes directly alter VS activation. The availability of an assay for gating currents in Hv1 may aid future efforts to elucidate the molecular mechanisms of gating cooperativity, pH-dependent modulation, and H%2b selectivity in a model VS domain protein. © 2018 Biophysical Society
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Hv1 channel, human; ion channel; cell line; channel gating; chemistry; genetics; human; kinetics; metabolism; mutation; pH; protein domain; thermodynamics; Cell Line; Humans; Hydrogen-Ion Concentration; Ion Channel Gating; Ion Channels; Kinetics; Mutation; Protein Domains; Thermodynamics
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