{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:17:06Z","timestamp":1792675026341,"version":"4.5.0"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2016,7,22]],"date-time":"2016-07-22T00:00:00Z","timestamp":1469145600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2016,7,22]],"date-time":"2016-07-22T00:00:00Z","timestamp":1469145600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The genome of <jats:italic>Weissella oryzae<\/jats:italic> SG25T was recently sequenced and a botulinum neurotoxin (BoNT) like gene was identified by bioinformatics methods. The typical three-domains organization of BoNTs with a N-terminal metalloprotease domain, a translocation and a cell binding domains could be identified. The BoNT family of neurotoxins is rapidly growing, but this was the first indication of the possible expression of a BoNT toxin outside the <jats:italic>Clostridium<\/jats:italic> genus. We performed molecular modeling and dynamics simulations showing that the 50\u2009kDa N-terminal domain folds very similarly to the metalloprotease domain of BoNT\/B, whilst the binding part is different. However, neither the recombinant metalloprotease nor the binding domains showed cross-reactivity with the standard antisera that define the seven serotypes of BoNTs. We found that the purified <jats:italic>Weissella<\/jats:italic> metalloprotease cleaves VAMP at a single site untouched by the other VAMP-specific BoNTs. This site is a unique Trp-Trp peptide bond located within the juxtamembrane segment of VAMP which is essential for neurotransmitter release. Therefore, the present study identifies the first non-Clostridial BoNT-like metalloprotease that cleaves VAMP at a novel and relevant site and we propose to label it BoNT\/Wo.<\/jats:p>","DOI":"10.1038\/srep30257","type":"journal-article","created":{"date-parts":[[2016,7,22]],"date-time":"2016-07-22T10:24:55Z","timestamp":1469183095000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":98,"title":["The first non Clostridial botulinum-like toxin cleaves VAMP within the juxtamembrane domain"],"prefix":"10.1038","volume":"6","author":[{"given":"Irene","family":"Zornetta","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Domenico","family":"Azarnia Tehran","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giorgio","family":"Arrigoni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabrizio","family":"Anniballi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Bano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oneda","family":"Leka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Zanotti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Binz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cesare","family":"Montecucco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,7,22]]},"reference":[{"key":"BFsrep30257_CR1","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1016\/S0072-9752(07)01511-4","volume":"91","author":"EA Johnson","year":"2008","unstructured":"Johnson, E. A. & Montecucco, C. Botulism. Handb Clin Neurol 91, 333\u2013368 (2008).","journal-title":"Handb Clin Neurol"},{"key":"BFsrep30257_CR2","first-page":"1","volume":"364","author":"KK Hill","year":"2013","unstructured":"Hill, K. K. & Smith, T. J. Genetic diversity within Clostridium botulinum serotypes, botulinum neurotoxin gene clusters and toxin subtypes. Curr Top Microbiol Immunol 364, 1\u201320 (2013).","journal-title":"Curr Top Microbiol Immunol"},{"key":"BFsrep30257_CR3","unstructured":"Centers for Disease Control and Prevention, D. o. H. a. H. S. Vol. 77 61083\u201361115 (Fed. Regist., 2012)."},{"key":"BFsrep30257_CR4","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1007\/s00018-013-1380-7","volume":"71","author":"S Pantano","year":"2014","unstructured":"Pantano, S. & Montecucco, C. The blockade of the neurotransmitter release apparatus by botulinum neurotoxins. Cell Mol Life Sci. 71, 793\u2013811 (2014).","journal-title":"Cell Mol Life Sci."},{"key":"BFsrep30257_CR5","doi-asserted-by":"crossref","unstructured":"S\u00fcdhof, T. C. & Rizo, J. Synaptic vesicle exocytosis. Cold Spring Harb Perspect Biol. 3 (2011).","DOI":"10.1101\/cshperspect.a005637"},{"key":"BFsrep30257_CR6","doi-asserted-by":"crossref","unstructured":"Rossetto, O., Pirazzini, M. & Montecucco, C. Botulinum neurotoxins: genetic, structural and mechanistic insights. Nat Rev Microbiol 12 (2014).","DOI":"10.1038\/nrmicro3295"},{"key":"BFsrep30257_CR7","doi-asserted-by":"publisher","first-page":"898","DOI":"10.1038\/2338","volume":"5","author":"DB Lacy","year":"1998","unstructured":"Lacy, D. B., Tepp, W., Cohen, A. C., DasGupta, B. R. & Stevens, R. C. Crystal structure of botulinum neurotoxin type A and implications for toxicity. Nat Struct Biol. 5, 898\u2013902 (1998).","journal-title":"Nat Struct Biol."},{"key":"BFsrep30257_CR8","doi-asserted-by":"publisher","first-page":"693","DOI":"10.1038\/78005","volume":"7","author":"S Swaminathan","year":"2000","unstructured":"Swaminathan, S. & Eswaramoorthy, S. Structural analysis of the catalytic and binding sites of Clostridium botulinum neurotoxin B. Nat Struct Biol. 7, 693\u2013699 (2000).","journal-title":"Nat Struct Biol."},{"key":"BFsrep30257_CR9","doi-asserted-by":"publisher","first-page":"591","DOI":"10.1146\/annurev.biochem.051908.125345","volume":"79","author":"M Montal","year":"2010","unstructured":"Montal, M. Botulinum neurotoxin: a marvel of protein design. Annu Rev Biochem 79, 591\u2013617 (2010).","journal-title":"Annu Rev Biochem"},{"key":"BFsrep30257_CR10","doi-asserted-by":"publisher","first-page":"665","DOI":"10.3390\/toxins2040665","volume":"2","author":"T Binz","year":"2010","unstructured":"Binz, T., Sikorra, S. & Mahrhold, S. Clostridial neurotoxins: mechanism of SNARE cleavage and outlook on potential substrate specificity reengineering. Toxins (Basel) 2, 665\u2013682 (2010).","journal-title":"Toxins (Basel)"},{"key":"BFsrep30257_CR11","doi-asserted-by":"publisher","first-page":"467","DOI":"10.1016\/j.bbamem.2015.08.014","volume":"1858","author":"M Pirazzini","year":"2016","unstructured":"Pirazzini, M. et al. On the translocation of botulinum and tetanus neurotoxins across the membrane of acidic intracellular compartments. Biochim Biophys Acta. 1858, 467\u2013474 (2016).","journal-title":"Biochim Biophys Acta."},{"key":"BFsrep30257_CR12","first-page":"61","volume":"364","author":"A Rummel","year":"2013","unstructured":"Rummel, A. Double receptor anchorage of botulinum neurotoxins accounts for their exquisite neurospecificity. Curr Top Microbiol Immunol 364, 61\u201390 (2013).","journal-title":"Curr Top Microbiol Immunol"},{"key":"BFsrep30257_CR13","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1016\/j.mib.2012.05.012","volume":"15","author":"D Dressler","year":"2012","unstructured":"Dressler, D. Clinical applications of botulinum toxin. Curr Opin Microbiol 15, 325\u2013336 (2012).","journal-title":"Curr Opin Microbiol"},{"key":"BFsrep30257_CR14","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.toxicon.2012.12.004","volume":"67","author":"M Hallett","year":"2013","unstructured":"Hallett, M. et al. Evidence-based review and assessment of botulinum neurotoxin for the treatment of movement disorders. Toxicon 67, 94\u2013114 (2013).","journal-title":"Toxicon"},{"key":"BFsrep30257_CR15","doi-asserted-by":"publisher","first-page":"628","DOI":"10.1002\/mus.20033","volume":"29","author":"CL Comella","year":"2004","unstructured":"Comella, C. L. & Pullman, S. L. Botulinum toxins in neurological disease. Muscle Nerve 29, 628\u2013644 (2004).","journal-title":"Muscle Nerve"},{"key":"BFsrep30257_CR16","doi-asserted-by":"publisher","first-page":"S53","DOI":"10.1002\/mds.20010","volume":"19","author":"R Eleopra","year":"2004","unstructured":"Eleopra, R., Tugnoli, V., Quatrale, R., Rossetto, O. & Montecucco, C. Different types of botulinum toxin in humans. Mov Disord 19 Suppl 8, S53\u2013S59 (2004).","journal-title":"Mov Disord"},{"key":"BFsrep30257_CR17","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1146\/annurev-pharmtox-011613-135935","volume":"54","author":"G Masuyer","year":"2014","unstructured":"Masuyer, G., Chaddock, J. A., Foster, K. A. & Acharya, K. R. Engineered botulinum neurotoxins as new therapeutics. Annu Rev Pharmacol Toxicol 54, 27\u201351 (2014).","journal-title":"Annu Rev Pharmacol Toxicol"},{"key":"BFsrep30257_CR18","doi-asserted-by":"crossref","unstructured":"Tanizawa, Y. et al. Draft Genome Sequence of Weissella oryzae SG25T, Isolated from Fermented Rice Grains. Genome Announc 2 (2014).","DOI":"10.1128\/genomeA.00667-14"},{"key":"BFsrep30257_CR19","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1111\/jfd.12236","volume":"38","author":"FA Costa","year":"2015","unstructured":"Costa, F. A., Leal, C. A., Schuenker, N. D., Leite, R. C. & Figueiredo, H. C. Characterization of Weissella ceti infections in Brazilian rainbow trout, Oncorhynchus mykiss (Walbaum), farms and development of an oil-adjuvanted vaccine. J Fish Dis. 38, 295\u2013302 (2015).","journal-title":"J Fish Dis."},{"key":"BFsrep30257_CR20","doi-asserted-by":"publisher","first-page":"254","DOI":"10.3389\/fmicb.2014.00254","volume":"5","author":"MR Fairfax","year":"2014","unstructured":"Fairfax, M. R., Lephart, P. R. & Salimnia, H. Weissella confusa: problems with identification of an opportunistic pathogen that has been found in fermented foods and proposed as a probiotic. Front Microbiol 5, 254, doi: 10.3389\/fmicb.2014.00254 (2014).","journal-title":"Front Microbiol"},{"key":"BFsrep30257_CR21","doi-asserted-by":"publisher","first-page":"155","DOI":"10.3389\/fmicb.2015.00155","volume":"6","author":"V Fusco","year":"2015","unstructured":"Fusco, V. et al. The genus Weissella: taxonomy, ecology and biotechnological potential. Front Microbiol 6, 155 (2015).","journal-title":"Front Microbiol"},{"key":"BFsrep30257_CR22","doi-asserted-by":"publisher","first-page":"342","DOI":"10.1016\/j.febslet.2014.12.018","volume":"589","author":"MJ Mansfield","year":"2015","unstructured":"Mansfield, M. J., Adams, J. B. & Doxey, A. C. Botulinum neurotoxin homologs in non-Clostridium species. FEBS Lett. 589, 342\u2013348 (2015).","journal-title":"FEBS Lett."},{"key":"BFsrep30257_CR23","doi-asserted-by":"publisher","first-page":"1096","DOI":"10.1038\/nature05411","volume":"444","author":"Q Chai","year":"2006","unstructured":"Chai, Q. et al. Structural basis of cell surface receptor recognition by botulinum neurotoxin B. Nature 444, 1096\u20131100 (2006).","journal-title":"Nature"},{"key":"BFsrep30257_CR24","doi-asserted-by":"publisher","first-page":"845","DOI":"10.1093\/bioinformatics\/btt055","volume":"29","author":"S Pronk","year":"2013","unstructured":"Pronk, S. et al. GROMACS 4.5: a high-throughput and highly parallel open source molecular simulation toolkit. Bioinformatics 29, 845\u2013854 (2013).","journal-title":"Bioinformatics"},{"key":"BFsrep30257_CR25","doi-asserted-by":"publisher","first-page":"431","DOI":"10.1016\/j.mam.2009.07.006","volume":"30","author":"F Tonello","year":"2009","unstructured":"Tonello, F. & Montecucco, C. The anthrax lethal factor and its MAPK kinase-specific metalloprotease activity. Mol Aspects Med. 30, 431\u2013438 (2009).","journal-title":"Mol Aspects Med."},{"key":"BFsrep30257_CR26","doi-asserted-by":"publisher","first-page":"1717","DOI":"10.1021\/bi0157969","volume":"41","author":"T Binz","year":"2002","unstructured":"Binz, T., Bade, S., Rummel, A., Kollewe, A. & Alves, J. Arg(362) and Tyr(365) of the botulinum neurotoxin type a light chain are involved in transition state stabilization. Biochemistry 41, 1717\u20131723 (2002).","journal-title":"Biochemistry"},{"key":"BFsrep30257_CR27","doi-asserted-by":"publisher","first-page":"1231","DOI":"10.1006\/bbrc.2001.5911","volume":"288","author":"M Rigoni","year":"2001","unstructured":"Rigoni, M., Caccin, P., Johnson, E. A., Montecucco, C. & Rossetto, O. Site-directed mutagenesis identifies active-site residues of the light chain of botulinum neurotoxin type A. Biochem Biophys Res Commun 288, 1231\u20131237 (2001).","journal-title":"Biochem Biophys Res Commun"},{"key":"BFsrep30257_CR28","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1093\/infdis\/jiv327","volume":"213","author":"SE Maslanka","year":"2016","unstructured":"Maslanka, S. E. et al. A Novel Botulinum Neurotoxin, Previously Reported as Serotype H, Has a Hybrid-Like Structure With Regions of Similarity to the Structures of Serotypes A and F and Is Neutralized With Serotype A Antitoxin. J Infect Dis. 213, 379\u2013385 (2016).","journal-title":"J Infect Dis."},{"key":"BFsrep30257_CR29","doi-asserted-by":"publisher","first-page":"643","DOI":"10.1016\/0076-6879(95)48041-2","volume":"248","author":"G Schiavo","year":"1995","unstructured":"Schiavo, G. & Montecucco, C. Tetanus and botulism neurotoxins: isolation and assay. Methods Enzymol 248, 643\u2013652 (1995).","journal-title":"Methods Enzymol"},{"key":"BFsrep30257_CR30","doi-asserted-by":"publisher","first-page":"8378","DOI":"10.1073\/pnas.0602644103","volume":"103","author":"M Bowen","year":"2006","unstructured":"Bowen, M. & Brunger, A. T. Conformation of the synaptobrevin transmembrane domain. Proc Natl Acad Sci USA 103, 8378\u20138383 (2006).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep30257_CR31","doi-asserted-by":"publisher","first-page":"959","DOI":"10.1016\/j.bpj.2012.08.007","volume":"103","author":"M Lindau","year":"2012","unstructured":"Lindau, M., Hall, B. A., Chetwynd, A., Beckstein, O. & Sansom, M. S. Coarse-grain simulations reveal movement of the synaptobrevin C-terminus in response to piconewton forces. Biophys J. 103, 959\u2013969 (2012).","journal-title":"Biophys J."},{"key":"BFsrep30257_CR32","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1016\/j.febslet.2012.11.002","volume":"587","author":"Q Fang","year":"2013","unstructured":"Fang, Q., Zhao, Y. & Lindau, M. Juxtamembrane tryptophans of synaptobrevin 2 control the process of membrane fusion. FEBS Lett. 587, 67\u201372 (2013).","journal-title":"FEBS Lett."},{"key":"BFsrep30257_CR33","doi-asserted-by":"publisher","first-page":"2994","DOI":"10.1016\/j.bbamem.2012.07.018","volume":"1818","author":"MS Al-Abdul-Wahid","year":"2012","unstructured":"Al-Abdul-Wahid, M. S., Demill, C. M., Serwin, M. B., Prosser, R. S. & Stewart, B. A. Effect of juxtamembrane tryptophans on the immersion depth of Synaptobrevin, an integral vesicle membrane protein. Biochim Biophys Acta 1818, 2994\u20132999 (2012).","journal-title":"Biochim Biophys Acta"},{"key":"BFsrep30257_CR34","doi-asserted-by":"publisher","first-page":"341","DOI":"10.1016\/0896-6273(95)90290-2","volume":"14","author":"ST Sweeney","year":"1995","unstructured":"Sweeney, S. T., Broadie, K., Keane, J., Niemann, H. & O\u2019Kane, C. J. Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects. Neuron 14, 341\u2013351 (1995).","journal-title":"Neuron"},{"key":"BFsrep30257_CR35","doi-asserted-by":"publisher","first-page":"682","DOI":"10.1016\/j.tibs.2004.10.002","volume":"29","author":"V Rossi","year":"2004","unstructured":"Rossi, V. et al. Longins and their longin domains: regulated SNAREs and multifunctional SNARE regulators. Trends Biochem Sci. 29, 682\u2013688 (2004).","journal-title":"Trends Biochem Sci."},{"key":"BFsrep30257_CR36","doi-asserted-by":"publisher","first-page":"e1003690","DOI":"10.1371\/journal.ppat.1003690","volume":"9","author":"K Lee","year":"2013","unstructured":"Lee, K. et al. Structure of a bimodular botulinum neurotoxin complex provides insights into its oral toxicity. PLoS Pathog 9, e1003690 (2013).","journal-title":"PLoS Pathog"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep30257","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep30257.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep30257.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,1,4]],"date-time":"2023-01-04T20:20:04Z","timestamp":1672863604000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep30257"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,7,22]]},"references-count":36,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2016,9,30]]}},"alternative-id":["BFsrep30257"],"URL":"https:\/\/doi.org\/10.1038\/srep30257","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,7,22]]},"assertion":[{"value":"23 May 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 July 2016","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 July 2016","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"30257"}}