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Botulinum Toxin in the Treatment of Headache
Open AccessArticle

Engineering an Effective Human SNAP-23 Cleaving Botulinum Neurotoxin A Variant

1
Institut für Zellbiochemie, OE 4310, Medizinische Hochschule Hannover, 30623 Hannover, Germany
2
Ipsen Bioinnovation, 102 Park Drive, Milton Park, Abingdon OX14 4RY, UK
*
Authors to whom correspondence should be addressed.
Toxins 2020, 12(12), 804; https://doi.org/10.3390/toxins12120804
Received: 17 November 2020 / Revised: 11 December 2020 / Accepted: 13 December 2020 / Published: 18 December 2020
(This article belongs to the Section Bacterial Toxins)
Botulinum neurotoxin (BoNT) serotype A inhibits neurotransmitter release by cleaving SNAP-25 and represents an established pharmaceutical for treating medical conditions caused by hyperactivity of cholinergic nerves. Oversecretion from non-neuronal cells is often also the cause of diseases. Notably, excessive release of inflammatory messengers is thought to contribute to diseases such as chronic obstructive pulmonary disease, asthma, diabetes etc. The expansion of its application to these medical conditions is prevented because the major non-neuronal SNAP-25 isoform responsible for exocytosis, SNAP-23, is, in humans, virtually resistant to BoNT/A. Based on previous structural data and mutagenesis studies of SNAP-23 we optimized substrate binding pockets of the enzymatic domain for interaction with SNAP-23. Systematic mutagenesis and rational design yielded the mutations E148Y, K166F, S254A, and G305D, each of which individually increased the activity of LC/A against SNAP-23 between 3- to 23-fold. The assembled quadruple mutant showed approximately 2000-fold increased catalytic activity against human SNAP-23 in in vitro cleavage assays. A comparable increase in activity was recorded for the full-length BoNT/A quadruple mutant tested in cultivated primary neurons transduced with a fluorescently tagged-SNAP-23 encoding gene. Equipped with a suitable targeting domain this quadruple mutant promises to complete successfully tests in cells of the immune system. View Full-Text
Keywords: SNAP-25; SNAP-23; zinc protease; botulinum toxin; substrate specificity; screening method SNAP-25; SNAP-23; zinc protease; botulinum toxin; substrate specificity; screening method
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MDPI and ACS Style

Sikorra, S.; Donald, S.; Elliott, M.; Schwede, S.; Coker, S.-F.; Kupinski, A.P.; Tripathi, V.; Foster, K.; Beard, M.; Binz, T. Engineering an Effective Human SNAP-23 Cleaving Botulinum Neurotoxin A Variant. Toxins 2020, 12, 804. https://doi.org/10.3390/toxins12120804

AMA Style

Sikorra S, Donald S, Elliott M, Schwede S, Coker S-F, Kupinski AP, Tripathi V, Foster K, Beard M, Binz T. Engineering an Effective Human SNAP-23 Cleaving Botulinum Neurotoxin A Variant. Toxins. 2020; 12(12):804. https://doi.org/10.3390/toxins12120804

Chicago/Turabian Style

Sikorra, Stefan; Donald, Sarah; Elliott, Mark; Schwede, Susan; Coker, Shu-Fen; Kupinski, Adam P.; Tripathi, Vineeta; Foster, Keith; Beard, Matthew; Binz, Thomas. 2020. "Engineering an Effective Human SNAP-23 Cleaving Botulinum Neurotoxin A Variant" Toxins 12, no. 12: 804. https://doi.org/10.3390/toxins12120804

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