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

Stefan Sikorra; Martin Skiba; Martin B. Dorner; Jasmin Weisemann; Mirjam Weil; Sylvia Valdezate; Bazbek Davletov; Andreas Rummel; Brigitte G. Dorner; Thomas Binz

doi:10.20944/preprints201806.0244.v1

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preprints.org > biology and life sciences > biochemistry and molecular biology > doi: 10.20944/preprints201806.0244.v1

Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

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

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Version 1 : Received: 14 June 2018 / Approved: 15 June 2018 / Online: 15 June 2018 (05:32:49 CEST)

A peer-reviewed article of this Preprint also exists.

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. 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.

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, but are also recognized pharmaceuticals for 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% on 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 factor eight. K_M values increase in the order F1 > F9 > F6 ≈ F7, whereas k_cat 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.

Keywords

Clostridium botulinum; botulinum neurotoxin; serotype F; subtype; VAMP-2; synaptobrevin; Zn2+ protease

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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