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Epitope Targeting of Tertiary Protein Structure Enables Target-Guided Synthesis of a Potent In-Cell Inhibitor of Botulinum Neurotoxin

Farrow, Blake and Wong, Michelle and Malette, Jacquie and Lai, Bert and Deyle, Kaycie M. and Das, Samir and Nag, Arundhati and Agnew, Heather D. and Heath, James R. (2015) Epitope Targeting of Tertiary Protein Structure Enables Target-Guided Synthesis of a Potent In-Cell Inhibitor of Botulinum Neurotoxin. Angewandte Chemie International Edition, 54 (24). pp. 7114-7119. ISSN 1433-7851. PMCID PMC5444873. https://resolver.caltech.edu/CaltechAUTHORS:20150504-105133177

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Abstract

Botulinum neurotoxin (BoNT) serotype A is the most lethal known toxin and has an occluded structure, which prevents direct inhibition of its active site before it enters the cytosol. Target-guided synthesis by in situ click chemistry is combined with synthetic epitope targeting to exploit the tertiary structure of the BoNT protein as a landscape for assembling a competitive inhibitor. A substrate-mimicking peptide macrocycle is used as a direct inhibitor of BoNT. An epitope-targeting in situ click screen is utilized to identify a second peptide macrocycle ligand that binds to an epitope that, in the folded BoNT structure, is active-site-adjacent. A second in situ click screen identifies a molecular bridge between the two macrocycles. The resulting divalent inhibitor exhibits an in vitro inhibition constant of 165 pM against the BoNT/A catalytic chain. The inhibitor is carried into cells by the intact holotoxin, and demonstrates protection and rescue of BoNT intoxication in a human neuron model.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/anie.201502451DOIArticle
https://onlinelibrary.wiley.com/doi/10.1002/anie.201502451/suppinfoPublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444873PubMed CentralArticle
ORCID:
AuthorORCID
Heath, James R.0000-0001-5356-4385
Additional Information:© 2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Received: March 16, 2015; Revised: April 9, 2015; Article first published online: 29 Apr. 2015. This work was supported by the Institute for Collaborative Biotechnologies (W911NF-09-0001) from the U.S. Army Research Office and the Defense Advanced Research Projects Agency (DARPA) through the Cooperative Agreement HR0011-11-2-0006, and the Jean Perkins Foundation. B.F. is supported by an HHMI International Student Research Fellowship. The following reagents were obtained through the NIH Biodefense and Emerging Infections Research Resources Repository, NIAID, NIH: Polyclonal anti- BoNT/A1 produced in sheep, NR-9584.
Funders:
Funding AgencyGrant Number
Army Research Office (ARO)W911NF-09-0001
Defense Advanced Research Projects Agency (DARPA)HR0011-11-2-0006
Jean Perkins FoundationUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
Subject Keywords:botulinum neurotoxin; combinatorial screening; epitope targeting; peptides; target-guided synthesis
Issue or Number:24
PubMed Central ID:PMC5444873
Record Number:CaltechAUTHORS:20150504-105133177
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150504-105133177
Official Citation:Farrow, B., Wong, M., Malette, J., Lai, B., Deyle, K. M., Das, S., Nag, A., Agnew, H. D. and Heath, J. R. (2015), Epitope Targeting of Tertiary Protein Structure Enables Target-Guided Synthesis of a Potent In-Cell Inhibitor of Botulinum Neurotoxin. Angew. Chem. Int. Ed., 54: 7114–7119. doi: 10.1002/anie.201502451
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:57191
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:04 May 2015 18:16
Last Modified:18 Dec 2019 23:58

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