Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells

Lam, Kwok-Ho; Perry, Kay; Shoemaker, Charles B.; Jin, Rongsheng

doi:10.3390/toxins12100616

Title: Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells

Journal Article · Sun Sep 27 00:00:00 EDT 2020 · Toxins

DOI:https://doi.org/10.3390/toxins12100616· OSTI ID:1673777

Lam, Kwok-Ho ^[1];

^[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)

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.

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Research Organization:: Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: USDOE; National Institutes of Health (NIH); National Institute of Allergy and Infections Diseases (NIAID)

Grant/Contract Number:: R01AI139087; R21AI139690; R21AI123920; R01AI125704; P30 GM124165; S10OD021527; AC02-06CH11357

OSTI ID:: 1673777

Journal Information:: Toxins, Vol. 12, Issue 10; ISSN 2072-6651

Publisher:: MDPICopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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60 APPLIED LIFE SCIENCES
botulinum neurotoxin
botulism
single-domain antibody
VHH
neutralizing epitope
antitoxin
membrane translocation

Title: Two VHH Antibodies Neutralize Botulinum Neurotoxin E1 by Blocking Its Membrane Translocation in Host Cells

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