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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2018
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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. |
format | Online Article Text |
id | pubmed-6304397 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
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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