{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,19]],"date-time":"2026-09-19T07:35:13Z","timestamp":1789803313525,"version":"4.0.1"},"reference-count":55,"publisher":"Rockefeller University Press","issue":"3","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2001,9,1]]},"abstract":"<jats:p>\u03b2-Scorpion toxins shift the voltage dependence of activation of sodium channels to more negative membrane potentials, but only after a strong depolarizing prepulse to fully activate the channels. Their receptor site includes the S3\u2013S4 loop at the extracellular end of the S4 voltage sensor in domain II of the \u03b1 subunit. Here, we probe the role of gating charges in the IIS4 segment in \u03b2-scorpion toxin action by mutagenesis and functional analysis of the resulting mutant sodium channels. Neutralization of the positively charged amino acid residues in the IIS4 segment by mutation to glutamine shifts the voltage dependence of channel activation to more positive membrane potentials and reduces the steepness of voltage-dependent gating, which is consistent with the presumed role of these residues as gating charges. Surprisingly, neutralization of the gating charges at the outer end of the IIS4 segment by the mutations R850Q, R850C, R853Q, and R853C markedly enhances \u03b2-scorpion toxin action, whereas mutations R856Q, K859Q, and K862Q have no effect. In contrast to wild-type, the \u03b2-scorpion toxin Css IV causes a negative shift of the voltage dependence of activation of mutants R853Q and R853C without a depolarizing prepulse at holding potentials from \u221280 to \u2212140 mV. Reaction of mutant R853C with 2-aminoethyl methanethiosulfonate causes a positive shift of the voltage dependence of activation and restores the requirement for a depolarizing prepulse for Css IV action. Enhancement of sodium channel activation by Css IV causes large tail currents upon repolarization, indicating slowed deactivation of the IIS4 voltage sensor by the bound toxin. Our results are consistent with a voltage-sensor\u2013trapping model in which the \u03b2-scorpion toxin traps the IIS4 voltage sensor in its activated position as it moves outward in response to depolarization and holds it there, slowing its inward movement on deactivation and enhancing subsequent channel activation. Evidently, neutralization of R850 and R853 removes kinetic barriers to binding of the IIS4 segment by Css IV, and thereby enhances toxin-induced channel activation.<\/jats:p>","DOI":"10.1085\/jgp.118.3.291","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T16:50:20Z","timestamp":1027702220000},"page":"291-302","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":65,"title":["Neutralization of Gating Charges in Domain II of the Sodium Channel \u03b1 Subunit Enhances Voltage-Sensor Trapping by a \u03b2-Scorpion Toxin"],"prefix":"10.1085","volume":"118","author":[{"given":"Sandrine","family":"Cest\u00e8le","sequence":"first","affiliation":[{"name":"aDepartment of Pharmacology, University of Washington School of Medicine, Seattle, WA 98195-7280"},{"name":"bUniversit\u00e9 de la M\u00e9diterran\u00e9e, I.F.R. Jean Roche, Centre National de la Recherche Scientifique, 13916 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Todd","family":"Scheuer","sequence":"additional","affiliation":[{"name":"aDepartment of Pharmacology, University of Washington School of Medicine, Seattle, WA 98195-7280"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Massimo","family":"Mantegazza","sequence":"additional","affiliation":[{"name":"aDepartment of Pharmacology, University of Washington School of Medicine, Seattle, WA 98195-7280"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herv\u00e9","family":"Rochat","sequence":"additional","affiliation":[{"name":"bUniversit\u00e9 de la M\u00e9diterran\u00e9e, I.F.R. Jean Roche, Centre National de la Recherche Scientifique, 13916 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William A.","family":"Catterall","sequence":"additional","affiliation":[{"name":"aDepartment of Pharmacology, University of Washington School of Medicine, Seattle, WA 98195-7280"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2001,8,27]]},"reference":[{"key":"2023072616522374800_Armstrong1981","doi-asserted-by":"crossref","first-page":"644","DOI":"10.1152\/physrev.1981.61.3.644","article-title":"Sodium channels and gating currents","volume":"61","author":"Armstrong","year":"1981","journal-title":"Physiol. 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