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Front. Microbiol. | doi: 10.3389/fmicb.2018.03083

Quantitative phosphoproteome analysis of Clostridioides difficile toxin B treated human epithelial cells

  • 1Hannover Medical School, Germany

The large clostridial glucosylating toxin B (TcdB) is a major virulence factor of the nosocomial pathogen Clostridioides difficile. TcdB inhibits small GTPases by glucosylation leading to impaired downstream signaling. TcdB also possesses a glucosyltransferase independent effect described as pyknosis. To elucidate the impact of TcdB and its glucosylation-inactive mutant TcdBNXN on the kinome of human cells, SILAC labeled HEp-2 cells were treated with 2 nM TcdB for 8 h. Phosphopeptides were enriched using SCX chromatography, IMAC and TiO2 followed shotgun mass spectrometry analysis. Overall 4,197 phosphopeptides were identified; more than 1,200 phosphosites responded to treatment with TcdB or TcdBNXN. The data suggested that predominantly stress-activated MAPK-dependent signaling pathways were triggered by toxin B treatment.

Keywords: Clostrioides difficile, Phophoproteomics, shot-gun proteomics, small GTPases, TCDB

Received: 15 Jun 2018; Accepted: 29 Nov 2018.

Edited by:

Miguel C. Teixeira, Universidade de Lisboa, Portugal

Reviewed by:

Philip R. Hardwidge, Kansas State University, United States
Raquel Hontecillas, Virginia Tech, United States  

Copyright: © 2018 Pich, Junemann, Just and Gerhard. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Prof. Andreas Pich, Hannover Medical School, Hanover, Germany, pich.andreas@mh-hannover.de