title: The first non Clostridial botulinum-like toxin cleaves VAMP within the juxtamembrane domain.
abstract: The genome of Weissella oryzae 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 Clostridium genus. We performed molecular modeling and dynamics simulations showing that the 50 kDa 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 Weissella 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.
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Zornetta
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Azarnia Tehran
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.|Proteomics Center, University of Padova and Azienda Ospedaliera di Padova, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Arrigoni
authors.affiliation: National Reference Center for Botulism, Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità (ISS), Roma, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Anniballi
authors.affiliation: Microbiology and Diagnostic Laboratory, Istituto Zooprofilattico Sperimentale delle Venezie (IZSVe), Vicolo Mazzini 4, Villorba di Treviso, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bano
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.
authors.forename: Oneda
authors.initials: O
authors.lastname: Leka
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Zanotti
authors.affiliation: Medizinische Hochschule Hannover, Institut für Physiologische Chemie OE4310, Hannover, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Binz
authors.affiliation: Department of Biomedical Sciences, University of Padova, Via Ugo Bassi 58/B, Padova, Italy.
authors.forename: Cesare
authors.initials: C
authors.lastname: Montecucco
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics; D001905:Botulinum Toxins / Q000737:chemistry* / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics; D003014:Clostridium botulinum / Q000235:genetics; D016680:Genome, Bacterial; D045726:Metalloproteases / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D056004:Molecular Dynamics Simulation; D009498:Neurotoxins / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D000072417:Protein Domains; D017510:Protein Folding; D058836:Weissella / Q000737:chemistry / Q000235:genetics*
chemical_list: D009498:Neurotoxins; D045726:Metalloproteases; D001905:Botulinum Toxins