TcdB of Clostridioides difficile Mediates RAS-Dependent Necrosis in Epithelial Cells
Abstract
:1. Introduction
2. Results
2.1. Morphological Changes after Treatment with Toxins
2.2. Comparative Phosphoproteomic Analysis
2.3. Kinetics of Phosphoproteome Alteration Point to RAS as Central Upstream Regulator
2.4. RAS Inhibition Led to a Reduction in Pyknosis
2.5. RAS Is Activated after TcdB Treatment
2.6. RAS Inhibition Influences Differences in Motif Phosphorylation and Reduced APOC3 Expression
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Protein Digestion
4.3. Phosphopeptide Enrichment and TMT Labeling
4.4. LC–MS Analysis
4.5. Flow Cytometry Analysis
4.6. DAPI Staining
4.7. RAS Activation Assay
4.8. Western Blotting
4.9. Data Processing
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Stieglitz, F.; Gerhard, R.; Hönig, R.; Giehl, K.; Pich, A. TcdB of Clostridioides difficile Mediates RAS-Dependent Necrosis in Epithelial Cells. Int. J. Mol. Sci. 2022, 23, 4258. https://doi.org/10.3390/ijms23084258
Stieglitz F, Gerhard R, Hönig R, Giehl K, Pich A. TcdB of Clostridioides difficile Mediates RAS-Dependent Necrosis in Epithelial Cells. International Journal of Molecular Sciences. 2022; 23(8):4258. https://doi.org/10.3390/ijms23084258
Chicago/Turabian StyleStieglitz, Florian, Ralf Gerhard, Rabea Hönig, Klaudia Giehl, and Andreas Pich. 2022. "TcdB of Clostridioides difficile Mediates RAS-Dependent Necrosis in Epithelial Cells" International Journal of Molecular Sciences 23, no. 8: 4258. https://doi.org/10.3390/ijms23084258