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Open AccessArticle

Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H

Ipsen Bioinnovation Ltd., 102 Park Drive, Milton Park, Abingdon OX14 4RY, UK
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Toxins 2018, 10(5), 195; https://doi.org/10.3390/toxins10050195
Received: 17 April 2018 / Revised: 4 May 2018 / Accepted: 8 May 2018 / Published: 11 May 2018
(This article belongs to the Special Issue Novel BoNTs and Toxin Engineering)
We have purified and characterized recombinant botulinum neurotoxin serotype FA (BoNT/FA). This protein has also been named as a new serotype (serotype H), but the classification has been controversial. A lack of well-characterized, highly pure material has been a roadblock to study. Here we report purification and characterization of enzymatically active, and of inactive nontoxic, recombinant forms of BoNT/FA as tractable alternatives to purifying this neurotoxin from native Clostridium botulinum. BoNT/FA cleaves the same intracellular target proteins as BoNT/F1 and other F serotype BoNTs; the intracellular targets are vesicle associated membrane proteins (VAMP) 1, 2 and 3. BoNT/FA cleaves the same site in VAMP-2 as BoNT/F5, which is different from the cleavage site of other F serotype BoNTs. BoNT/FA has slower enzyme kinetics than BoNT/F1 in a cell-free protease assay and is less potent at inhibiting ex vivo nerve-stimulated skeletal muscle contraction. In contrast, BoNT/FA is more potent at inhibiting neurotransmitter release from cultured neurons. View Full-Text
Keywords: bacterial toxin; botulinum toxin; neurotoxin; neurotransmitter release; protein purification; proteolytic enzyme; recombinant protein expression; SNARE proteins bacterial toxin; botulinum toxin; neurotoxin; neurotransmitter release; protein purification; proteolytic enzyme; recombinant protein expression; SNARE proteins
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MDPI and ACS Style

Hackett, G.; Moore, K.; Burgin, D.; Hornby, F.; Gray, B.; Elliott, M.; Mir, I.; Beard, M. Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H. Toxins 2018, 10, 195. https://doi.org/10.3390/toxins10050195

AMA Style

Hackett G, Moore K, Burgin D, Hornby F, Gray B, Elliott M, Mir I, Beard M. Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H. Toxins. 2018; 10(5):195. https://doi.org/10.3390/toxins10050195

Chicago/Turabian Style

Hackett, Gavin; Moore, Kevin; Burgin, David; Hornby, Fraser; Gray, Bryony; Elliott, Mark; Mir, Imran; Beard, Matthew. 2018. "Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H" Toxins 10, no. 5: 195. https://doi.org/10.3390/toxins10050195

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