{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T23:46:55Z","timestamp":1784677615813,"version":"3.55.0"},"reference-count":34,"publisher":"Portland Press Ltd.","issue":"7","license":[{"start":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T00:00:00Z","timestamp":1721088000000},"content-version":"vor","delay-in-days":15,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003074","name":"Agencia Nacional de Promoci\u00f3n Cient\u00edfica y Tecnol\u00f3gica","doi-asserted-by":"crossref","award":["PICT2020-02021"],"award-info":[{"award-number":["PICT2020-02021"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100003074","name":"Agencia Nacional de Promoci\u00f3n Cient\u00edfica y Tecnol\u00f3gica","doi-asserted-by":"crossref","award":["PICT2019-01889"],"award-info":[{"award-number":["PICT2019-01889"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100020302","name":"Secretar\u00eda de Investigaci\u00f3n, Internacionales y Posgrado, Universidad Nacional de Cuyo","doi-asserted-by":"crossref","award":["06\/M033-T1"],"award-info":[{"award-number":["06\/M033-T1"]}],"id":[{"id":"10.13039\/501100020302","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2024,7,31]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Botulinum neurotoxins (BoNTs) are valuable tools to unveil molecular mechanisms of exocytosis in neuronal and non-neuronal cells due to their peptidase activity on exocytic isoforms of SNARE proteins. They are produced by Clostridia as single-chain polypeptides that are proteolytically cleaved into light, catalytic domains covalently linked via disulfide bonds to heavy, targeting domains. This format of two subunits linked by disulfide bonds is required for the full neurotoxicity of BoNTs. We have generated a recombinant version of BoNT\/B that consists of the light chain of the toxin fused to the protein transduction domain of the human immunodeficiency virus-1 (TAT peptide) and a hexahistidine tag. His6-TAT-BoNT\/B-LC, expressed in Escherichia coli and purified by affinity chromatography, penetrated membranes and exhibited strong enzymatic activity, as evidenced by cleavage of the SNARE synaptobrevin from rat brain synaptosomes and human sperm cells. Proteolytic attack of synaptobrevin hindered exocytosis triggered by a calcium ionophore in the latter. The novel tool reported herein disrupts the function of a SNARE protein within minutes in cells that may or may not express the receptors for the BoNT\/B heavy chain, and without the need for transient transfection or permeabilization.<\/jats:p>","DOI":"10.1042\/bsr20240117","type":"journal-article","created":{"date-parts":[[2024,7,16]],"date-time":"2024-07-16T08:27:31Z","timestamp":1721118451000},"update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Expression, purification and application of a recombinant, membrane permeating version of the light chain of botulinum toxin B"],"prefix":"10.1042","volume":"44","author":[{"given":"Micaela\u00a0Vanina","family":"Buzzatto","sequence":"first","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabiana\u00a0Cristina","family":"Benegas Guerrero","sequence":"additional","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pablo\u00a0Ariel","family":"\u00c1lvarez","sequence":"additional","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mar\u00eda\u00a0Paz","family":"Zizzias","sequence":"additional","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis\u00a0Mariano","family":"Polo","sequence":"additional","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3376-6868","authenticated-orcid":false,"given":"Claudia\u00a0Nora","family":"Tomes","sequence":"additional","affiliation":[{"name":"1Instituto de Histolog\u00eda y Embriolog\u00eda de Mendoza (IHEM)-CONICET-Universidad Nacional de Cuyo, Argentina"},{"name":"2Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Cuyo, Argentina"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2024,7,30]]},"reference":[{"key":"2024073006364391300_B1","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1038\/s41580-023-00668-x","article-title":"Mechanisms of SNARE proteins in membrane fusion","volume":"25","author":"Jahn","year":"2024","journal-title":"Nat. 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