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Botulinum Neurotoxin Diversity from a Gene-Centered View
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Toxins 2018, 10(8), 311; https://doi.org/10.3390/toxins10080311

Botulinum Neurotoxin F Subtypes Cleaving the VAMP-2 Q58–K59 Peptide Bond Exhibit Unique Catalytic Properties and Substrate Specificities

1
Institute of Cell Biochemistry, OE 4310, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany
2
Biological Toxins (ZBS 3), Centre for Biological Threats and Special Pathogens, Robert Koch Institute, 13353 Berlin, Germany
3
Institute of Toxicology, OE 5340, Hannover Medical School, Carl-Neuberg-Straße 1, 30625 Hannover, Germany
4
Reference and Research Laboratory for Taxonomy, Spanish National Centre of Microbiology, Institute of Health Carlos III, 28220 Madrid, Spain
5
Department of Biomedical Science, University of Sheffield, Western Bank, Sheffield S10 2TN, UK
*
Authors to whom correspondence should be addressed.
Received: 14 June 2018 / Revised: 23 July 2018 / Accepted: 30 July 2018 / Published: 1 August 2018
(This article belongs to the Special Issue Novel BoNTs and Toxin Engineering)
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Abstract

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 > F7 > F9 > F6, and vary by up to a factor of eight. KM values increase in the order F1 > F9 > F6 ≈ F7, whereas kcat decreases in the order F7 > F1 > F9 > 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. View Full-Text
Keywords: Clostridium botulinum; botulinum neurotoxin; serotype F; subtype; vesicle associated membrane protein 2 (VAMP-2); synaptobrevin; Zn2+ protease Clostridium botulinum; botulinum neurotoxin; serotype F; subtype; vesicle associated membrane protein 2 (VAMP-2); synaptobrevin; Zn2+ protease
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Sikorra, S.; Skiba, M.; Dorner, M.B.; Weisemann, J.; Weil, M.; Valdezate, S.; Davletov, B.; Rummel, A.; Dorner, B.G.; Binz, T. Botulinum Neurotoxin F Subtypes Cleaving the VAMP-2 Q58–K59 Peptide Bond Exhibit Unique Catalytic Properties and Substrate Specificities. Toxins 2018, 10, 311.

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