{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T12:24:04Z","timestamp":1793276644195,"version":"4.5.0"},"reference-count":50,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,1,21]],"date-time":"2022-01-21T00:00:00Z","timestamp":1642723200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003032","name":"Association Nationale de la Recherche et de la Technologie","doi-asserted-by":"publisher","award":["525\/2016"],"award-info":[{"award-number":["525\/2016"]}],"id":[{"id":"10.13039\/501100003032","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-11-LABX-0015"],"award-info":[{"award-number":["ANR-11-LABX-0015"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001665","name":"Agence Nationale de la Recherche","doi-asserted-by":"publisher","award":["ANR-18--CE19-0024-02"],"award-info":[{"award-number":["ANR-18--CE19-0024-02"]}],"id":[{"id":"10.13039\/501100001665","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Conseil R\u00e9gional de Pays de la Loire","award":["2016-11092\/11093"],"award-info":[{"award-number":["2016-11092\/11093"]}]},{"DOI":"10.13039\/501100001674","name":"Fondation Leducq","doi-asserted-by":"publisher","award":["Equipement de recherche et plateforme technologique"],"award-info":[{"award-number":["Equipement de recherche et plateforme technologique"]}],"id":[{"id":"10.13039\/501100001674","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002924","name":"FEDER","doi-asserted-by":"publisher","award":["2017 \/ FEDER \/ PL0014592"],"award-info":[{"award-number":["2017 \/ FEDER \/ PL0014592"]}],"id":[{"id":"10.13039\/501100002924","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>\u03b1-bungarotoxin is a large, 74 amino acid toxin containing five disulphide bridges, initially identified in the venom of Bungarus multicinctus snake. Like most large toxins, chemical synthesis of \u03b1-bungarotoxin is challenging, explaining why all previous reports use purified or recombinant \u03b1-bungarotoxin. However, only chemical synthesis allows easy insertion of non-natural amino acids or new chemical functionalities. Herein, we describe a procedure for the chemical synthesis of a fluorescent-tagged \u03b1-bungarotoxin. The full-length peptide was designed to include an alkyne function at the amino-terminus through the addition of a pentynoic acid linker. Chemical synthesis of \u03b1-bungarotoxin requires hydrazide-based coupling of three peptide fragments in successive steps. After completion of the oxidative folding, an azide-modified Cy5 fluorophore was coupled by click chemistry onto the toxin. Next, we determined the efficacy of the fluorescent-tagged \u03b1-bungarotoxin to block acetylcholine (ACh)-mediated currents in response to muscle nicotinic receptor activation in TE671 cells. Using automated patch-clamp recordings, we demonstrate that fluorescent synthetic \u03b1-bungarotoxin has the expected nanomolar affinity for the nicotinic receptor. The blocking effect of fluorescent \u03b1-bungarotoxin could be displaced by incubation with a 20-mer peptide mimicking the \u03b1-bungarotoxin binding site. In addition, TE671 cells could be labelled with fluorescent toxin, as witnessed by confocal microscopy, and this labelling was partially displaced by the 20-mer competitive peptide. We thus demonstrate that synthetic fluorescent-tagged \u03b1-bungarotoxin preserves excellent properties for binding onto muscle nicotinic receptors.<\/jats:p>","DOI":"10.3390\/toxins14020079","type":"journal-article","created":{"date-parts":[[2022,1,23]],"date-time":"2022-01-23T20:36:27Z","timestamp":1642970187000},"page":"79","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Chemical Synthesis of a Functional Fluorescent-Tagged \u03b1-Bungarotoxin"],"prefix":"10.3390","volume":"14","author":[{"given":"Oliver","family":"Brun","sequence":"first","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"},{"name":"Faculty of Medicine, Universit\u00e9 de Montr\u00e9al, Montr\u00e9al, QC H3T 1J4, Canada"},{"name":"Research Center, Montr\u00e9al Heart Institute, Montr\u00e9al, QC H1T 1C8, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claude","family":"Zoukimian","sequence":"additional","affiliation":[{"name":"Smartox Biotechnology, 6 Rue des Platanes, F-38120 Saint-Egr\u00e8ve, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2269-5750","authenticated-orcid":false,"given":"Barbara","family":"Oliveira-Mendes","sequence":"additional","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5100-5571","authenticated-orcid":false,"given":"J\u00e9r\u00f4me","family":"Montnach","sequence":"additional","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1370-6155","authenticated-orcid":false,"given":"Benjamin","family":"Lauzier","sequence":"additional","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michel","family":"Ronjat","sequence":"additional","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R\u00e9my","family":"B\u00e9roud","sequence":"additional","affiliation":[{"name":"Smartox Biotechnology, 6 Rue des Platanes, F-38120 Saint-Egr\u00e8ve, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3699-1283","authenticated-orcid":false,"given":"Fr\u00e9d\u00e9ric","family":"Lesage","sequence":"additional","affiliation":[{"name":"Research Center, Montr\u00e9al Heart Institute, Montr\u00e9al, QC H1T 1C8, Canada"},{"name":"\u00c9cole Polytechnique Montr\u00e9al, Montreal, QC H3C 3A7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2530-0299","authenticated-orcid":false,"given":"Didier","family":"Boturyn","sequence":"additional","affiliation":[{"name":"UMR 5250, D\u00e9partement de Chimie Mol\u00e9culaire, CNRS, Universit\u00e9 Grenoble Alpes, CEDEX 09, F-38058 Grenoble, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2782-9615","authenticated-orcid":false,"given":"Michel","family":"De Waard","sequence":"additional","affiliation":[{"name":"L\u2019institut Du Thorax, Nantes Universit\u00e9, INSERM, CNRS, INSERM UMR 1087\/CNRS UMR 6291, 8 Quai Moncousu, F-44007 Nantes, France"},{"name":"Smartox Biotechnology, 6 Rue des Platanes, F-38120 Saint-Egr\u00e8ve, France"},{"name":"LabEx \u201cIon Channels, Science & Therapeutics\u201d, F-06560 Valbonne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1515\/bchm2.1972.353.1.243","article-title":"Purification, properties and amino acid sequence of alpha-bungarotoxin from the venom of Bungarus multicinctus","volume":"353","author":"Mebs","year":"1972","journal-title":"Hoppe-Seyler\u2019s Z. 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