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Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells

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 pyknosi...

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Autores principales: Junemann, Johannes, Just, Ingo, Gerhard, Ralf, Pich, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6304397/
https://www.ncbi.nlm.nih.gov/pubmed/30619164
http://dx.doi.org/10.3389/fmicb.2018.03083
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author Junemann, Johannes
Just, Ingo
Gerhard, Ralf
Pich, Andreas
author_facet Junemann, Johannes
Just, Ingo
Gerhard, Ralf
Pich, Andreas
author_sort Junemann, Johannes
collection PubMed
description 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 TcdB(NXN) 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 TiO(2) followed shotgun mass spectrometry analysis. Overall 4,197 phosphopeptides were identified; more than 1,200 phosphosites responded to treatment with TcdB or TcdB(NXN). The data suggested that predominantly stress-activated MAPK-dependent signaling pathways were triggered by toxin B treatment.
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spelling pubmed-63043972019-01-07 Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells Junemann, Johannes Just, Ingo Gerhard, Ralf Pich, Andreas Front Microbiol Microbiology 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 TcdB(NXN) 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 TiO(2) followed shotgun mass spectrometry analysis. Overall 4,197 phosphopeptides were identified; more than 1,200 phosphosites responded to treatment with TcdB or TcdB(NXN). The data suggested that predominantly stress-activated MAPK-dependent signaling pathways were triggered by toxin B treatment. Frontiers Media S.A. 2018-12-17 /pmc/articles/PMC6304397/ /pubmed/30619164 http://dx.doi.org/10.3389/fmicb.2018.03083 Text en Copyright © 2018 Junemann, Just, Gerhard and Pich. http://creativecommons.org/licenses/by/4.0/ 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.
spellingShingle Microbiology
Junemann, Johannes
Just, Ingo
Gerhard, Ralf
Pich, Andreas
Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title_full Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title_fullStr Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title_full_unstemmed Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title_short Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells
title_sort quantitative phosphoproteome analysis of clostridioides difficile toxin b treated human epithelial cells
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6304397/
https://www.ncbi.nlm.nih.gov/pubmed/30619164
http://dx.doi.org/10.3389/fmicb.2018.03083
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