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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-12-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2018.03083/full |
| _version_ | 1819260230322618368 |
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| author | Johannes Junemann Ingo Just Ralf Gerhard Andreas Pich |
| author_facet | Johannes Junemann Ingo Just Ralf Gerhard Andreas Pich |
| author_sort | Johannes Junemann |
| collection | DOAJ |
| 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 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. |
| first_indexed | 2024-12-23T19:22:36Z |
| format | Article |
| id | doaj.art-644d01d4e0ab4730b627371bc507d559 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-23T19:22:36Z |
| publishDate | 2018-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-644d01d4e0ab4730b627371bc507d5592022-12-21T17:34:06ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-12-01910.3389/fmicb.2018.03083408346Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial CellsJohannes JunemannIngo JustRalf GerhardAndreas PichThe 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.https://www.frontiersin.org/article/10.3389/fmicb.2018.03083/fullClostridioides difficilephosphoproteomeshotgun proteomicssmall GTPasesTcdB |
| spellingShingle | Johannes Junemann Ingo Just Ralf Gerhard Andreas Pich Quantitative Phosphoproteome Analysis of Clostridioides difficile Toxin B Treated Human Epithelial Cells Frontiers in Microbiology Clostridioides difficile phosphoproteome shotgun proteomics small GTPases TcdB |
| 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 | Clostridioides difficile phosphoproteome shotgun proteomics small GTPases TcdB |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2018.03083/full |
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