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Jingzhaotoxin\u2010III (\u03b2\u2010TRTX\u2010Cj1\u03b1) is a unique sodium channel gating modifier from the tarantula\n                      <jats:italic>Chilobrachys jingzhao<\/jats:italic>\n                      , because the toxin can selectively inhibit the activation of cardiac sodium channel but not neuronal subtypes. However, the molecular basis of JZTX\u2010III interaction with sodium channels remains unknown. In this study, we showed that JZTX\u2010III was efficiently expressed by the secretory pathway in yeast. Alanine\u2010scanning analysis indicated that 2 acidic residues (Asp1, Glu3) and an exposed hydrophobic patch, formed by 4 Trp residues (residues 8, 9, 28 and 30), play important roles in the binding of JZTX\u2010III to Nav1.5. JZTX\u2010III docked to the Nav1.5 DIIS3\u2010S4 linker. Mutations S799A, R800A, and L804A could additively reduce toxin sensitivity of Nav1.5. We also demonstrated that the unique Arg800, not emerging in other sodium channel subtypes, is responsible for JZTX\u2010III selectively interacting with Nav1.5. The reverse mutation D816R in Nav1.7 greatly increased the sensitivity of the neuronal subtype to JZTX\u2010III. Conversely, the mutation R800D in Nav1.5 decreased JZTX\u2010III's IC\n                      <jats:sub>50<\/jats:sub>\n                      by 72\u2010fold. Therefore, our results indicated that JZTX\u2010III is a site 4 toxin, but does not possess the same critical residues on sodium channels as other site 4 toxins. Our data also revealed the underlying mechanism for JZTX\u2010III to be highly specific for the cardiac sodium channel.\u2014Rong, M., Chen, J., Tao, H., Wu, Y., Jiang, P., Lu, M., Su, H., Chi, Y., Cai, T., Zhao, L., Zeng, X., Xiao, Y., Liang, S. Molecular basis of the tarantula toxin jingzhaotoxin\u2010III (\u03b2\u2010TRTX\u2010Cj1\u03b1) interacting with voltage sensors in sodium channel subtype Nav1.5.\n                      <jats:italic>FASEB J.<\/jats:italic>\n                      25, 3177\u20103185 (2011).\n                      <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.fasebj.org\">www.fasebj.org<\/jats:ext-link>\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1096\/fj.10-178848","type":"journal-article","created":{"date-parts":[[2011,6,10]],"date-time":"2011-06-10T23:09:35Z","timestamp":1307747375000},"page":"3177-3185","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Molecular basis of the tarantula toxin jingzhaotoxin\u2010III (\u03b2\u2010TRTX\u2010Cj1\u03b1) interacting with voltage sensors in sodium channel subtype 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