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Toxin A of the nosocomial pathogen Clostridium difficile induces primary effects in the proteome of HEp‐2 cells
Author(s) -
Junemann Johannes,
Birgin Gurbet,
Erdmann Jelena,
Schröder Anke,
Just Ingo,
Gerhard Ralf,
Pich Andreas
Publication year - 2017
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201600031
Subject(s) - clostridium difficile , proteome , toxin , microbiology and biotechnology , clostridium difficile toxin a , pathogen , primary (astronomy) , biology , antibiotics , bioinformatics , physics , astronomy
Purpose This study was carried out to investigate the impact of high concentrations of Clostridium difficile toxin A (TcdA) on the proteome of human cells. It should also be examined whether a catalytically deficient mutant (TcdA NXN ) has an effect on target cells. Experimental design Proteome changes were investigated after treatment of HEp‐2 cells with 20 nM TcdA for 8 h using a triplex SILAC labeling method and shotgun proteomics. Proteins from differently labeled and treated cells were combined for analysis using an HPLC coupled to an Orbitrap mass spectrometer. Results Nearly 4000 proteins were identified in each replicate and 3500 could be quantified by SILAC triplicate analysis. 51 proteins exhibited an altered abundance with 29 up‐regulated and 22 down‐regulated proteins. In contrast, TcdA NXN had no provable impact on the protein profile of HEp‐2 cells. Data analysis of regulated proteins revealed that mainly plasma membrane, cell death, cell proliferation and actin cytoskeleton proteins were affected by TcdA treatment. Conclusions and clinical relevance This proteome analysis showed novel insights of TcdA impact onepithelial cells. Comparison with long‐term treatment studies reveals distinctions in affected cellular processes that will improve the understanding of TcdA functions and might help to find new tools for diagnosis and treatment of CDI.