{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T01:53:56Z","timestamp":1783130036053,"version":"3.54.6"},"reference-count":68,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2023,8,12]],"date-time":"2023-08-12T00:00:00Z","timestamp":1691798400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"University of Bath"},{"name":"Ipsen Bioinnovation Ltd."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Clostridium botulinum neurotoxins (BoNTs) are the most potent toxins known, causing the deadly disease botulism. They function through Zn2+-dependent endopeptidase cleavage of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins, preventing vesicular fusion and subsequent neurotransmitter release from motor neurons. Several serotypes of BoNTs produced by Clostridium botulinum (BoNT\/A-\/G and\/X) have been well-characterised over the years. However, a BoNT-like gene (homologue of BoNT) was recently identified in the non-clostridial species, Enterococcus faecium, which is the leading cause of hospital-acquired multi-drug resistant infections. Here, we report the crystal structure of the catalytic domain of a BoNT homologue from Enterococcus faecium (LC\/En) at 2.0 \u00c5 resolution. Detailed structural analysis in comparison with the full-length BoNT\/En AlphaFold2-predicted structure, LC\/A (from BoNT\/A), and LC\/F (from BoNT\/F) revealed putative subsites and exosites (including loops 1\u20135) involved in recognition of LC\/En substrates. LC\/En also appears to possess a conserved autoproteolytic cleavage site whose function is yet to be established.<\/jats:p>","DOI":"10.3390\/ijms241612721","type":"journal-article","created":{"date-parts":[[2023,8,14]],"date-time":"2023-08-14T10:13:57Z","timestamp":1692008037000},"page":"12721","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Crystal Structure of the Catalytic Domain of a Botulinum Neurotoxin Homologue from Enterococcus faecium: Potential Insights into Substrate Recognition"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5371-4283","authenticated-orcid":false,"given":"Kyle S.","family":"Gregory","sequence":"first","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6832-0922","authenticated-orcid":false,"given":"Peter-Rory","family":"Hall","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jude Prince","family":"Onuh","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Otsile O.","family":"Mojanaga","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6359-0229","authenticated-orcid":false,"given":"Sai Man","family":"Liu","sequence":"additional","affiliation":[{"name":"Protein Sciences Department, Ipsen Bioinnovation Limited, 102 Park Drive, Milton Park, Abingdon OX14 4RY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3009-4058","authenticated-orcid":false,"given":"K. Ravi","family":"Acharya","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, University of Bath, Claverton Down, Bath BA2 7AY, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e1065","DOI":"10.7717\/peerj.1065","article-title":"The Destructive Effect of Botulinum Neurotoxins on the SNARE Protein: SNAP-25 and Synaptic Membrane Fusion","volume":"3","author":"Lu","year":"2015","journal-title":"PeerJ"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1007\/164_2019_342","article-title":"The Structure and Classification of Botulinum Toxins","volume":"263","author":"Dong","year":"2021","journal-title":"Botulinum Toxin Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1146\/annurev-biochem-013118-111654","article-title":"Botulinum and Tetanus Neurotoxins","volume":"88","author":"Dong","year":"2019","journal-title":"Annu. 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