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Later experiments revealed that GrTX-SIA could also inhibit voltage-gated K+ (KV) channel currents via a similar mechanism of action that involved binding to a conserved S3\u2013S4 region in the voltage-sensing domains (VSDs). Since voltage-gated Na+ (NaV) channels contain homologous structural motifs, we hypothesized that GrTX-SIA could inhibit members of this ion channel family as well. Here, we show that GrTX-SIA can indeed impede the gating process of multiple NaV channel subtypes with NaV1.6 being the most susceptible target. Moreover, molecular docking of GrTX-SIA onto NaV1.6, supported by a p.E1607K mutation, revealed the voltage sensor in domain IV (VSDIV) as being a primary site of action. The biphasic manner in which current inhibition appeared to occur suggested a second, possibly lower-sensitivity binding locus, which was identified as VSDII by using KV2.1\/NaV1.6 chimeric voltage-sensor constructs. Subsequently, the NaV1.6p.E782K\/p.E838K (VSDII), NaV1.6p.E1607K (VSDIV), and particularly the combined VSDII\/VSDIV mutant lost virtually all susceptibility to GrTX-SIA. Together with existing literature, our data suggest that GrTX-SIA recognizes modules in NaV channel VSDs that are conserved among ion channel families, thereby allowing it to act as a comprehensive ion channel gating modifier peptide.<\/jats:p>","DOI":"10.1085\/jgp.202413563","type":"journal-article","created":{"date-parts":[[2024,7,11]],"date-time":"2024-07-11T02:20:33Z","timestamp":1720664433000},"update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":2,"title":["\u03c9-Grammotoxin-SIA inhibits voltage-gated Na+ channel currents"],"prefix":"10.1085","volume":"156","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2826-1603","authenticated-orcid":true,"given":"Rita de C\u00e1ssia","family":"Colla\u00e7o","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Molecular Physiology and Neurophysics Group, Ghent University 1 Department of Basic and Applied Medical Sciences, , Ghent, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2728-8537","authenticated-orcid":true,"given":"Filip","family":"Van Petegem","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03rmrcq20","id-type":"ROR","asserted-by":"publisher"}],"name":"University of British Columbia 2 Department of Biochemistry and Molecular Biology, , Vancouver, Canada"},{"id":[{"id":"https:\/\/ror.org\/03rmrcq20","id-type":"ROR","asserted-by":"publisher"}],"name":"Life Sciences Institute, University of British Columbia 3 , Vancouver, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0451-6050","authenticated-orcid":true,"given":"Frank","family":"Bosmans","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00cv9y106","id-type":"ROR","asserted-by":"publisher"}],"name":"Molecular Physiology and Neurophysics Group, Ghent University 1 Department of Basic and Applied Medical Sciences, , Ghent, Belgium"},{"name":"Faculty of Medicine and Pharmacy, Center for Neurosciences (C4N), Vrije Universiteit Brussel 4 , Brussels, Belgium"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"2024072313493020100_bib1","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1085\/jgp.201511492","article-title":"The hitchhiker\u2019s guide to the voltage-gated sodium channel galaxy","volume":"147","author":"Ahern","year":"2016","journal-title":"J. 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