{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T07:05:21Z","timestamp":1783494321473,"version":"3.55.0"},"reference-count":35,"publisher":"Rockefeller University Press","issue":"2","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2014,8,1]]},"abstract":"<jats:p>Voltage-gated sodium channels mediate the initiation and propagation of action potentials in excitable cells. Transmembrane segment S4 of voltage-gated sodium channels resides in a gating pore where it senses the membrane potential and controls channel gating. Substitution of individual S4 arginine gating charges (R1\u2013R3) with smaller amino acids allows ionic currents to flow through the mutant gating pore, and these gating pore currents are pathogenic in some skeletal muscle periodic paralysis syndromes. The voltage dependence of gating pore currents provides information about the transmembrane position of the gating charges as S4 moves in response to membrane potential. Here we studied gating pore current in mutants of the homotetrameric bacterial sodium channel NaChBac in which individual arginine gating charges were replaced by cysteine. Gating pore current was observed for each mutant channel, but with different voltage-dependent properties. Mutating the first (R1C) or second (R2C) arginine to cysteine resulted in gating pore current at hyperpolarized membrane potentials, where the channels are in resting states, but not at depolarized potentials, where the channels are activated. Conversely, the R3C gating pore is closed at hyperpolarized membrane potentials and opens with channel activation. Negative conditioning pulses revealed time-dependent deactivation of the R3C gating pore at the most hyperpolarized potentials. Our results show sequential voltage dependence of activation of gating pore current from R1 to R3 and support stepwise outward movement of the substituted cysteines through the narrow portion of the gating pore that is sealed by the arginine side chains in the wild-type channel. This pattern of voltage dependence of gating pore current is consistent with a sliding movement of the S4 helix through the gating pore. Through comparison with high-resolution models of the voltage sensor of bacterial sodium channels, these results shed light on the structural basis for pathogenic gating pore currents in periodic paralysis syndromes.<\/jats:p>","DOI":"10.1085\/jgp.201411210","type":"journal-article","created":{"date-parts":[[2014,7,28]],"date-time":"2014-07-28T13:25:02Z","timestamp":1406553902000},"page":"147-157","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":32,"title":["Tracking S4 movement by gating pore currents in the bacterial sodium channel NaChBac"],"prefix":"10.1085","volume":"144","author":[{"given":"Tamer M.","family":"Gamal El-Din","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Todd","family":"Scheuer","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William A.","family":"Catterall","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Washington, Seattle, WA 98195"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2014,7,28]]},"reference":[{"key":"2023072623222754400_bib1","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1152\/physrev.2000.80.2.555","article-title":"The voltage sensor in voltage-dependent ion channels","volume":"80","author":"Bezanilla","year":"2000","journal-title":"Physiol. Rev."},{"key":"2023072623222754400_bib2","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1007\/s00232-007-9016-1","article-title":"Accessibility of four arginine residues on the S4 segment of the Bacillus halodurans sodium channel","volume":"215","author":"Blanchet","year":"2007","journal-title":"J. Membr. Biol."},{"key":"2023072623222754400_bib3","doi-asserted-by":"publisher","first-page":"2648","DOI":"10.1073\/pnas.1115575109","article-title":"Gating transitions in the selectivity filter region of a sodium channel are coupled to the domain IV voltage sensor","volume":"109","author":"Capes","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib4","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/S0896-6273(00)81133-2","article-title":"From ionic currents to molecular mechanisms: the structure and function of voltage-gated sodium channels","volume":"26","author":"Catterall","year":"2000","journal-title":"Neuron."},{"key":"2023072623222754400_bib5","doi-asserted-by":"publisher","first-page":"915","DOI":"10.1016\/j.neuron.2010.08.021","article-title":"Ion channel voltage sensors: structure, function, and pathophysiology","volume":"67","author":"Catterall","year":"2010","journal-title":"Neuron."},{"key":"2023072623222754400_bib6","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1007\/s00232-004-0701-z","article-title":"Role of arginine residues on the S4 segment of the Bacillus halodurans Na+ channel in voltage-sensing","volume":"201","author":"Chahine","year":"2004","journal-title":"J. Membr. Biol."},{"key":"2023072623222754400_bib7","doi-asserted-by":"publisher","first-page":"5435","DOI":"10.1073\/pnas.0914109107","article-title":"The activated state of a sodium channel voltage sensor in a membrane environment","volume":"107","author":"Chakrapani","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib8","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/BF00257102","article-title":"Quantal charge redistributions accompanying the structural transitions of sodium channels","volume":"17","author":"Conti","year":"1989","journal-title":"Eur. Biophys. J."},{"key":"2023072623222754400_bib9","doi-asserted-by":"publisher","first-page":"15142","DOI":"10.1073\/pnas.0806486105","article-title":"Disulfide locking a sodium channel voltage sensor reveals ion pair formation during activation","volume":"105","author":"DeCaen","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib10","doi-asserted-by":"publisher","first-page":"22498","DOI":"10.1073\/pnas.0912307106","article-title":"Sequential formation of ion pairs during activation of a sodium channel voltage sensor","volume":"106","author":"DeCaen","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib11","doi-asserted-by":"publisher","first-page":"18825","DOI":"10.1073\/pnas.1116449108","article-title":"Gating charge interactions with the S1 segment during activation of a Na+ channel voltage sensor","volume":"108","author":"DeCaen","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib12","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1085\/jgp.79.1.41","article-title":"Distribution and kinetics of membrane dielectric polarization. II. Frequency domain studies of gating currents","volume":"79","author":"Fern\u00e1ndez","year":"1982","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib13","doi-asserted-by":"publisher","first-page":"1165","DOI":"10.1529\/biophysj.108.130765","article-title":"More gating charges are needed to open a Shaker K+ channel than are needed to open an rBIIA Na+ channel","volume":"95","author":"Gamal El-Din","year":"2008","journal-title":"Biophys. J."},{"key":"2023072623222754400_bib14","doi-asserted-by":"publisher","first-page":"93","DOI":"10.4161\/chan.4.2.10672","article-title":"Double gaps along Shaker S4 demonstrate omega currents at three different closed states","volume":"4","author":"Gamal El-Din","year":"2010","journal-title":"Channels (Austin)."},{"key":"2023072623222754400_bib15","article-title":"Ionic channels of excitable membranes","author":"Hille","year":"2001","edition":"Third edition"},{"key":"2023072623222754400_bib16","doi-asserted-by":"publisher","first-page":"1053","DOI":"10.1085\/jgp.106.6.1053","article-title":"Transfer of twelve charges is needed to open skeletal muscle Na+ channels","volume":"106","author":"Hirschberg","year":"1995","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib17","doi-asserted-by":"publisher","first-page":"349","DOI":"10.1085\/jgp.200409139","article-title":"Gating of the bacterial sodium channel, NaChBac: Voltage-dependent charge movement and gating currents","volume":"124","author":"Kuzmenkin","year":"2004","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib18","doi-asserted-by":"publisher","first-page":"635","DOI":"10.1085\/jgp.201210827","article-title":"Intermediate state trapping of a voltage sensor","volume":"140","author":"Lacroix","year":"2012","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib19","doi-asserted-by":"publisher","first-page":"897","DOI":"10.1126\/science.1116269","article-title":"Crystal structure of a mammalian voltage-dependent Shaker family K+ channel","volume":"309","author":"Long","year":"2005","journal-title":"Science."},{"key":"2023072623222754400_bib20","doi-asserted-by":"publisher","first-page":"376","DOI":"10.1038\/nature06265","article-title":"Atomic structure of a voltage-dependent K+ channel in a lipid membrane-like environment","volume":"450","author":"Long","year":"2007","journal-title":"Nature."},{"key":"2023072623222754400_bib21","doi-asserted-by":"publisher","first-page":"353","DOI":"10.1038\/nature10238","article-title":"The crystal structure of a voltage-gated sodium channel","volume":"475","author":"Payandeh","year":"2011","journal-title":"Nature."},{"key":"2023072623222754400_bib22","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1038\/nature11077","article-title":"Crystal structure of a voltage-gated sodium channel in two potentially inactivated states","volume":"486","author":"Payandeh","year":"2012","journal-title":"Nature."},{"key":"2023072623222754400_bib23","doi-asserted-by":"publisher","first-page":"2372","DOI":"10.1126\/science.1065635","article-title":"A prokaryotic voltage-gated sodium channel","volume":"294","author":"Ren","year":"2001","journal-title":"Science."},{"key":"2023072623222754400_bib24","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/j.neuron.2005.06.012","article-title":"Ion permeation through a voltage-sensitive gating pore in brain sodium channels having voltage sensor mutations","volume":"47","author":"Sokolov","year":"2005","journal-title":"Neuron."},{"key":"2023072623222754400_bib25","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1038\/nature05598","article-title":"Gating pore current in an inherited ion channelopathy","volume":"446","author":"Sokolov","year":"2007","journal-title":"Nature."},{"key":"2023072623222754400_bib26","doi-asserted-by":"publisher","first-page":"19980","DOI":"10.1073\/pnas.0810562105","article-title":"Depolarization-activated gating pore current conducted by mutant sodium channels in potassium-sensitive normokalemic periodic paralysis","volume":"105","author":"Sokolov","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib27","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1085\/jgp.201010414","article-title":"Ion permeation and block of the gating pore in the voltage sensor of NaV1.4 channels with hypokalemic periodic paralysis mutations","volume":"136","author":"Sokolov","year":"2010","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib28","doi-asserted-by":"publisher","first-page":"548","DOI":"10.1038\/nature02270","article-title":"A proton pore in a potassium channel voltage sensor reveals a focused electric field","volume":"427","author":"Starace","year":"2004","journal-title":"Nature."},{"key":"2023072623222754400_bib29","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1085\/jgp.200709755","article-title":"A Na+ channel mutation linked to hypokalemic periodic paralysis exposes a proton-selective gating pore","volume":"130","author":"Struyk","year":"2007","journal-title":"J. Gen. Physiol."},{"key":"2023072623222754400_bib30","doi-asserted-by":"publisher","first-page":"597","DOI":"10.1038\/339597a0","article-title":"Structural parts involved in activation and inactivation of the sodium channel","volume":"339","author":"St\u00fchmer","year":"1989","journal-title":"Nature."},{"key":"2023072623222754400_bib31","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1016\/j.neuron.2004.12.047","article-title":"Voltage-sensing arginines in a potassium channel permeate and occlude cation-selective pores","volume":"45","author":"Tombola","year":"2005","journal-title":"Neuron."},{"key":"2023072623222754400_bib32","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/0896-6273(95)90078-0","article-title":"Evidence for voltage-dependent S4 movement in sodium channels","volume":"15","author":"Yang","year":"1995","journal-title":"Neuron."},{"key":"2023072623222754400_bib33","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1016\/S0896-6273(00)80028-8","article-title":"Molecular basis of charge movement in voltage-gated sodium channels","volume":"16","author":"Yang","year":"1996","journal-title":"Neuron."},{"key":"2023072623222754400_bib34","doi-asserted-by":"publisher","first-page":"E93","DOI":"10.1073\/pnas.1118434109","article-title":"Structural basis for gating charge movement in the voltage sensor of a sodium channel","volume":"109","author":"Yarov-Yarovoy","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072623222754400_bib35","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1038\/nature11054","article-title":"Crystal structure of an orthologue of the NaChBac voltage-gated sodium channel","volume":"486","author":"Zhang","year":"2012","journal-title":"Nature."}],"container-title":["Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/144\/2\/147\/1793570\/jgp_201411210.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/144\/2\/147\/1793570\/jgp_201411210.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,26]],"date-time":"2023-07-26T23:22:47Z","timestamp":1690413767000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/144\/2\/147\/53253\/Tracking-S4-movement-by-gating-pore-currents-in"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,7,28]]},"references-count":35,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2014,8,1]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.201411210","relation":{},"ISSN":["1540-7748","0022-1295"],"issn-type":[{"value":"1540-7748","type":"electronic"},{"value":"0022-1295","type":"print"}],"subject":[],"published":{"date-parts":[[2014,7,28]]}}}