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标题：Hydrophobic gasket mutation produces gating pore currents in closed human voltage-gated proton channels
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作者：Richard Banh ; Vladimir V. Cherny ; Deri Morgan 等
期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2019
卷号：116
期号：38
页码：18951-18961
DOI：10.1073/pnas.1905462116
出版社：The National Academy of Sciences of the United States of America
摘要：The hydrophobic gasket (HG), a ring of hydrophobic amino acids in the voltage-sensing domain of most voltage-gated ion channels, forms a constriction between internal and external aqueous vestibules. Cationic Arg or Lys side chains lining the S4 helix move through this “gating pore” when the channel opens. S4 movement may occur during gating of the human voltage-gated proton channel, hH_V1, but proton current flows through the same pore in open channels. Here, we replaced putative HG residues with less hydrophobic residues or acidic Asp. Substitution of individuals, pairs, or all 3 HG positions did not impair proton selectivity. Evidently, the HG does not act as a secondary selectivity filter. However, 2 unexpected functions of the HG in H_V1 were discovered. Mutating HG residues independently accelerated channel opening and compromised the closed state. Mutants exhibited open–closed gating, but strikingly, at negative voltages where “normal” gating produces a nonconducting closed state, the channel leaked protons. Closed-channel proton current was smaller than open-channel current and was inhibited by 10 μM Zn²⁺. Extreme hyperpolarization produced a deeper closed state through a weakly voltage-dependent transition. We functionally identify the HG as Val¹⁰⁹, Phe¹⁵⁰, Val¹⁷⁷, and Val¹⁷⁸, which play a critical and exclusive role in preventing H⁺ influx through closed channels. Molecular dynamics simulations revealed enhanced mobility of Arg²⁰⁸ in mutants exhibiting H⁺ leak. Mutation of HG residues produces gating pore currents reminiscent of several channelopathies.
关键词：HVCN1 ; voltage-sensing domain ; voltage gating ; ion channels ; protons