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These toxins recognise motor nerve terminals with high specificity and affinity by using a dual binding mechanism involving gangliosides and protein receptors. The initial recognition of gangliosides is crucial for the toxins\u2019 potency. In this study, we employed a synaptosome-binding screening strategy to identify BoNT\/A mutants with enhanced ganglioside-binding which translated into improved potency. X-ray crystallography and receptor-binding assays were used to elucidate the molecular mechanisms underlying the increased affinity or altered ganglioside selectivity of these mutants. Our findings provide a basis for the development of BoNT\/A variants with enhanced therapeutic potential.<\/jats:p>","DOI":"10.1038\/s42004-025-01569-0","type":"journal-article","created":{"date-parts":[[2025,6,4]],"date-time":"2025-06-04T16:02:00Z","timestamp":1749052920000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Botulinum neurotoxin A mutants with enhanced ganglioside binding show improved potency and altered ganglioside selectivity"],"prefix":"10.1038","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9527-2310","authenticated-orcid":false,"given":"Geoffrey","family":"Masuyer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0645-1641","authenticated-orcid":false,"given":"Andreas","family":"Rummel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4777-3417","authenticated-orcid":false,"given":"P\u00e5l","family":"Stenmark","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,6,4]]},"reference":[{"key":"1569_CR1","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1152\/physrev.2000.80.2.717","volume":"80","author":"G Schiavo","year":"2000","unstructured":"Schiavo, G., Matteoli, M. & Montecucco, C. 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A.R. is sole inventor of the granted patent family EP1786832, US8293230, etc. The authors declare no other competing interest.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"171"}}