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

Toxins
Gavin HackettMatthew Beard

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.

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Citations

Feb 29, 2020·Mini Reviews in Medicinal Chemistry·Miroslav Pohanka
Jul 14, 2018·Toxins·Alexander P Gardner, Joseph T Barbieri
Jan 8, 2021·Science Translational Medicine·Patrick M McNuttKonstantin Ichtchenko
Oct 12, 2021·Pharmacology Research & Perspectives·Cindy PérierMikhail Kalinichev

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Methods Mentioned

BETA
genetic modification
X-ray
Assay

Software Mentioned

GraphPad Prism
Genetools
GraphPad

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Botulism (ASM)

Botulism is a rare but serious paralytic illness caused by a nerve toxin that is produced by the bacterium clostridium botulinum. Discover the latest research on botulism here.

Botulism

Botulism is a rare but serious paralytic illness caused by a nerve toxin that is produced by the bacterium clostridium botulinum. Discover the latest research on botulism here.