Benjamin Spiller
Last active: 4/10/2019

A neutralizing antibody that blocks delivery of the enzymatic cargo of toxin TcdB into host cells.

Kroh HK, Chandrasekaran R, Zhang Z, Rosenthal K, Woods R, Jin X, Nyborg AC, Rainey GJ, Warrener P, Melnyk RA, Spiller BW, Lacy DB
J Biol Chem. 2018 293 (3): 941-952

PMID: 29180448 · PMCID: PMC5777265 · DOI:10.1074/jbc.M117.813428

infection is the leading cause of hospital-acquired diarrhea and is mediated by the actions of two toxins, TcdA and TcdB. The toxins perturb host cell function through a multistep process of receptor binding, endocytosis, low pH-induced pore formation, and the translocation and delivery of an N-terminal glucosyltransferase domain that inactivates host GTPases. Infection studies with isogenic strains having defined toxin deletions have established TcdB as an important target for therapeutic development. Monoclonal antibodies that neutralize TcdB function have been shown to protect against infection in animal models and reduce recurrence in humans. Here, we report the mechanism of TcdB neutralization by PA41, a humanized monoclonal antibody capable of neutralizing TcdB from a diverse array of strains. Through a combination of structural, biochemical, and cell functional studies, involving X-ray crystallography and EM, we show that PA41 recognizes a single, highly conserved epitope on the TcdB glucosyltransferase domain and blocks productive translocation and delivery of the enzymatic cargo into the host cell. Our study reveals a unique mechanism of toxin neutralization by a monoclonal antibody, which involves targeting a process that is conserved across the large clostridial glucosylating toxins. The PA41 antibody described here provides a valuable tool for dissecting the mechanism of toxin pore formation and translocation across the endosomal membrane.

MeSH Terms (13)

Antibodies, Monoclonal Antibodies, Neutralizing Bacterial Toxins Caco-2 Cells Clostridium difficile Crystallography, X-Ray Cytosol Enterotoxins Humans Hydrogen-Ion Concentration Microscopy, Electron rac1 GTP-Binding Protein Rubidium

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