Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells
Abstract
Botulinum neurotoxin serotype E (BoNT/E) is one of the major causes of human botulism, which is a life-threatening disease caused by flaccid paralysis of muscles. After receptor-mediated toxin internalization into motor neurons, the translocation domain (HN) of BoNT/E transforms into a protein channel upon vesicle acidification in endosomes and delivers its protease domain (LC) across membrane to enter the neuronal cytosol. It is believed that the rapid onset of BoNT/E intoxication compared to other BoNT serotypes is related to its swift internalization and translocation. We recently identified two neutralizing single-domain camelid antibodies (VHHs) against BoNT/E1 termed JLE-E5 and JLE-E9. Here, we report the crystal structures of these two VHHs bound to the LCHN domain of BoNT/E1. The structures reveal that these VHHs recognize two distinct epitopes that are partially overlapping with the putative transmembrane regions on HN, and therefore could physically block membrane association of BoNT/E1. This is confirmed by our in vitro studies, which show that these VHHs inhibit the structural change of BoNT/E1 at acidic pH and interfere with BoNT/E1 association with lipid vesicles. Therefore, these two VHHs neutralize BoNT/E1 by preventing the transmembrane delivery of LC. Furthermore, structure-based sequence analyses show that the 3-dimensional epitopes of thesemore »
- Authors:
-
[2];
[3];
[1]
- Univ. of California, Irvine, CA (United States)
- Cornell Univ., Ithaca, NY (United States); Argonne National Lab. (ANL), Argonne, IL (United States)
- Tufts Univ., North Grafton, MA (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
- Sponsoring Org.:
- USDOE; National Institutes of Health (NIH); National Institute of Allergy and Infections Diseases (NIAID)
- OSTI Identifier:
- 1673777
- Grant/Contract Number:
- R01AI139087; R21AI139690; R21AI123920; R01AI125704; P30 GM124165; S10OD021527; AC02-06CH11357
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Toxins
- Additional Journal Information:
- Journal Volume: 12; Journal Issue: 10; Journal ID: ISSN 2072-6651
- Publisher:
- MDPI
- Country of Publication:
- United States
- Language:
- ENGLISH
- Subject:
- 60 APPLIED LIFE SCIENCES; botulinum neurotoxin; botulism; single-domain antibody; VHH; neutralizing epitope; antitoxin; membrane translocation
Citation Formats
Lam, Kwok-Ho, Perry, Kay, Shoemaker, Charles B., and Jin, Rongsheng. Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells. United States: N. p., 2020.
Web. doi:10.3390/toxins12100616.
Lam, Kwok-Ho, Perry, Kay, Shoemaker, Charles B., & Jin, Rongsheng. Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells. United States. https://doi.org/10.3390/toxins12100616
Lam, Kwok-Ho, Perry, Kay, Shoemaker, Charles B., and Jin, Rongsheng. Sun .
"Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells". United States. https://doi.org/10.3390/toxins12100616. https://www.osti.gov/servlets/purl/1673777.
@article{osti_1673777,
title = {Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells},
author = {Lam, Kwok-Ho and Perry, Kay and Shoemaker, Charles B. and Jin, Rongsheng},
abstractNote = {Botulinum neurotoxin serotype E (BoNT/E) is one of the major causes of human botulism, which is a life-threatening disease caused by flaccid paralysis of muscles. After receptor-mediated toxin internalization into motor neurons, the translocation domain (HN) of BoNT/E transforms into a protein channel upon vesicle acidification in endosomes and delivers its protease domain (LC) across membrane to enter the neuronal cytosol. It is believed that the rapid onset of BoNT/E intoxication compared to other BoNT serotypes is related to its swift internalization and translocation. We recently identified two neutralizing single-domain camelid antibodies (VHHs) against BoNT/E1 termed JLE-E5 and JLE-E9. Here, we report the crystal structures of these two VHHs bound to the LCHN domain of BoNT/E1. The structures reveal that these VHHs recognize two distinct epitopes that are partially overlapping with the putative transmembrane regions on HN, and therefore could physically block membrane association of BoNT/E1. This is confirmed by our in vitro studies, which show that these VHHs inhibit the structural change of BoNT/E1 at acidic pH and interfere with BoNT/E1 association with lipid vesicles. Therefore, these two VHHs neutralize BoNT/E1 by preventing the transmembrane delivery of LC. Furthermore, structure-based sequence analyses show that the 3-dimensional epitopes of these two VHHs are largely conserved across many BoNT/E subtypes, suggesting a broad-spectrum protection against the BoNT/E family. In summary, this work improves our understanding of the membrane translocation mechanism of BoNT/E and paves the way for developing VHHs as diagnostics or therapeutics for the treatment of BoNT/E intoxication.},
doi = {10.3390/toxins12100616},
journal = {Toxins},
number = 10,
volume = 12,
place = {United States},
year = {Sun Sep 27 00:00:00 EDT 2020},
month = {Sun Sep 27 00:00:00 EDT 2020}
}
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