{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T16:41:25Z","timestamp":1791996085601,"version":"4.4.0"},"reference-count":45,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2018,8,1]],"date-time":"2018-08-01T00:00:00Z","timestamp":1533081600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["Bi 660\/3-1"],"award-info":[{"award-number":["Bi 660\/3-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Swiss Federal Department of Defence, Civil Protection and Sport","award":["353003364\/Stm"],"award-info":[{"award-number":["353003364\/Stm"]}]},{"DOI":"10.13039\/501100002347","name":"Federal Ministry of Education and Research","doi-asserted-by":"publisher","award":["031L0111A"],"award-info":[{"award-number":["031L0111A"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>In the recent past, about 40 botulinum neurotoxin (BoNT) subtypes belonging to serotypes A, B, E, and F pathogenic to humans were identified among hundreds of independent isolates. BoNTs are the etiological factors of botulism and represent potential bioweapons; however, they are also recognized pharmaceuticals for the efficient counteraction of hyperactive nerve terminals in a variety of human diseases. The detailed biochemical characterization of subtypes as the basis for development of suitable countermeasures and possible novel therapeutic applications is lagging behind the increase in new subtypes. Here, we report the primary structure of a ninth subtype of BoNT\/F. Its amino-acid sequence diverges by at least 8.4% at the holotoxin and 13.4% at the enzymatic domain level from all other known BoNT\/F subtypes. We found that BoNT\/F9 shares the scissile Q58\/K59 bond in its substrate vesicle associated membrane protein 2 with the prototype BoNT\/F1. Comparative biochemical analyses of four BoNT\/F enzymatic domains showed that the catalytic efficiencies decrease in the order F1 &gt; F7 &gt; F9 &gt; F6, and vary by up to a factor of eight. KM values increase in the order F1 &gt; F9 &gt; F6 \u2248 F7, whereas kcat decreases in the order F7 &gt; F1 &gt; F9 &gt; F6. Comparative substrate scanning mutagenesis studies revealed a unique pattern of crucial substrate residues for each subtype. Based upon structural coordinates of F1 bound to an inhibitor polypeptide, the mutational analyses suggest different substrate interactions in the substrate binding channel of each subtype.<\/jats:p>","DOI":"10.3390\/toxins10080311","type":"journal-article","created":{"date-parts":[[2018,8,1]],"date-time":"2018-08-01T11:22:34Z","timestamp":1533122554000},"page":"311","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Botulinum Neurotoxin F Subtypes Cleaving the VAMP-2 Q58\u2013K59 Peptide Bond Exhibit Unique Catalytic Properties and Substrate Specificities"],"prefix":"10.3390","volume":"10","author":[{"given":"Stefan","family":"Sikorra","sequence":"first","affiliation":[{"name":"Institute of Cell Biochemistry, OE 4310, Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Skiba","sequence":"additional","affiliation":[{"name":"Biological Toxins (ZBS 3), Centre for Biological Threats and Special Pathogens, Robert Koch Institute, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8176-797X","authenticated-orcid":false,"given":"Martin B.","family":"Dorner","sequence":"additional","affiliation":[{"name":"Biological Toxins (ZBS 3), Centre for Biological Threats and Special Pathogens, Robert Koch Institute, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6299-9257","authenticated-orcid":false,"given":"Jasmin","family":"Weisemann","sequence":"additional","affiliation":[{"name":"Institute of Toxicology, OE 5340, Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mirjam","family":"Weil","sequence":"additional","affiliation":[{"name":"Biological Toxins (ZBS 3), Centre for Biological Threats and Special Pathogens, Robert Koch Institute, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3931-2162","authenticated-orcid":false,"given":"Sylvia","family":"Valdezate","sequence":"additional","affiliation":[{"name":"Reference and Research Laboratory for Taxonomy, Spanish National Centre of Microbiology, Institute of Health Carlos III, 28220 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bazbek","family":"Davletov","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield S10 2TN, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0645-1641","authenticated-orcid":false,"given":"Andreas","family":"Rummel","sequence":"additional","affiliation":[{"name":"Institute of Toxicology, OE 5340, Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5100-2546","authenticated-orcid":false,"given":"Brigitte G.","family":"Dorner","sequence":"additional","affiliation":[{"name":"Biological Toxins (ZBS 3), Centre for Biological Threats and Special Pathogens, Robert Koch Institute, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9133-3617","authenticated-orcid":false,"given":"Thomas","family":"Binz","sequence":"additional","affiliation":[{"name":"Institute of Cell Biochemistry, OE 4310, Hannover Medical School, Carl-Neuberg-Stra\u00dfe 1, 30625 Hannover, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,8,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1002\/1873-3468.12969","article-title":"Identification of a novel botulinum neurotoxin gene cluster in Enterococcus","volume":"592","author":"Brunt","year":"2018","journal-title":"FEBS Lett."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/j.chom.2017.12.018","article-title":"Identification of a botulinum neurotoxin-like toxin in a commensal strain of Enterococcus faecium","volume":"23","author":"Zhang","year":"2018","journal-title":"Cell Host Microbe"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"14130","DOI":"10.1038\/ncomms14130","article-title":"Identification and characterization of a novel botulinum neurotoxin","volume":"8","author":"Zhang","year":"2017","journal-title":"Nat. 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