Toxins 2018, 10(5), 195; https://doi.org/10.3390/toxins10050195
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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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)
Abstract
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-TextKeywords:
bacterial toxin; botulinum toxin; neurotoxin; neurotransmitter release; protein purification; proteolytic enzyme; recombinant protein expression; SNARE proteins
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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.
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