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We aimed at producing ProTx II analogues entitled with novel functionalities for cell distribution studies and biochemical characterization of its Na<jats:sub>v<\/jats:sub> channel targets.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Approach<\/jats:title><jats:p>We took advantage of the high affinity properties of the peptide, combined to its slow off rate, to design a number of new tagged analogues useful for imaging and biochemistry purposes. We used high\u2010throughput automated patch\u2010clamp to identify the analogues best matching the native properties of ProTx II and validated them on various Na<jats:sub>v<\/jats:sub>\u2010expressing cells in pull\u2010down and cell distribution studies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>Two of the produced ProTx II analogues, Biot\u2010ProTx II and ATTO488\u2010ProTx II, best emulate the pharmacological properties of unlabelled ProTx II, whereas other analogues remain high affinity blockers of Na<jats:sub>v<\/jats:sub>1.7. The biotinylated version of ProTx II efficiently works for the pull\u2010down of several Na<jats:sub>v<\/jats:sub> isoforms tested in a concentration\u2010dependent manner, whereas the fluorescent ATTO488\u2010ProTx II specifically labels the Na<jats:sub>v<\/jats:sub>1.7 channel over other Na<jats:sub>v<\/jats:sub> isoforms tested in various experimental conditions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Implications<\/jats:title><jats:p>The properties of these ProTx II analogues as tools for Na<jats:sub>v<\/jats:sub> channel purification and cell distribution studies pave the way for a better understanding of ProTx II channel receptors in pain and their pathophysiological implications in sensory neuronal processing. The new fluorescent ProTx II should also be useful in the design of new drug screening strategies.<\/jats:p><\/jats:sec>","DOI":"10.1111\/bph.15453","type":"journal-article","created":{"date-parts":[[2021,3,20]],"date-time":"2021-03-20T05:05:34Z","timestamp":1616216734000},"page":"2632-2650","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Fluorescent\u2010 and tagged\u2010protoxin II peptides: potent markers of the Na<sub>v<\/sub>1.7 channel pain target"],"prefix":"10.1111","volume":"178","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5100-5571","authenticated-orcid":false,"given":"J\u00e9r\u00f4me","family":"Montnach","sequence":"first","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4236-6687","authenticated-orcid":false,"given":"Stephan","family":"De Waard","sequence":"additional","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S\u00e9bastien","family":"Nicolas","sequence":"additional","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0557-3396","authenticated-orcid":false,"given":"Sophie","family":"Burel","sequence":"additional","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nancy","family":"Osorio","sequence":"additional","affiliation":[{"name":"Laboratory of Cognitive Neuroscience, UMR 7291, CNRS Aix\u2010Marseille University  Marseille France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claude","family":"Zoukimian","sequence":"additional","affiliation":[{"name":"Smartox Biotechnology  Saint\u2010Egr\u00e8ve France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1070-7929","authenticated-orcid":false,"given":"Massimo","family":"Mantegazza","sequence":"additional","affiliation":[{"name":"Universit\u00e9 Cote d'Azur, CNRS UMR 7275 Institute of Molecular and Cellular Pharmacology  Valbonne\u2010Sophia Antipolis France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rachid","family":"Boukaiba","sequence":"additional","affiliation":[{"name":"Sanofi R&amp;D, Integrated Drug Discovery \u2013 High Content Biology  Vitry\u2010sur\u2010Seine France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R\u00e9my","family":"B\u00e9roud","sequence":"additional","affiliation":[{"name":"Smartox Biotechnology  Saint\u2010Egr\u00e8ve France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michel","family":"Partiseti","sequence":"additional","affiliation":[{"name":"Sanofi R&amp;D, Integrated Drug Discovery \u2013 High Content Biology  Vitry\u2010sur\u2010Seine France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0123-5413","authenticated-orcid":false,"given":"Patrick","family":"Delmas","sequence":"additional","affiliation":[{"name":"Laboratory of Cognitive Neuroscience, UMR 7291, CNRS Aix\u2010Marseille University  Marseille France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0962-9756","authenticated-orcid":false,"given":"C\u00e9line","family":"Marionneau","sequence":"additional","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2782-9615","authenticated-orcid":false,"given":"Michel","family":"De Waard","sequence":"additional","affiliation":[{"name":"LabEx \u201cIon Channels, Science &amp; Therapeutics\u201d l'institut du thorax, INSERM, CNRS, UNIV NANTES  Nantes France"},{"name":"Smartox Biotechnology  Saint\u2010Egr\u00e8ve France"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,5,14]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aax4755"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/bph.14749"},{"key":"e_1_2_12_4_1","doi-asserted-by":"crossref","unstructured":"Bilimoria P. 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