Quantitative Proteomic Analysis of the Heat Stress Response in Clostridium difficile Strain 630
Abstract
Clostridium difficile is a serious nosocomial pathogen whose prevalence worldwide is increasing. Postgenomic technologies can now be deployed to develop understanding of the evolution and diversity of this important human pathogen, yet little is known about the adaptive ability of C. difficile. We used iTRAQ labeling and 2D-LC–MS/MS driven proteomics to investigate the response of C. difficile 630 to a mild, but clinically relevant, heat stress. A statistically validated list of 447 proteins to which functional roles were assigned was generated, allowing reconstruction of central metabolic pathways including glycolysis, γ-aminobutyrate metabolism, and peptidoglycan biosynthesis. Some 49 proteins were significantly modulated under heat stress: classical heat shock proteins including GroEL, GroES, DnaK, Clp proteases, and HtpG were up-regulated in addition to several stress inducible rubrerythrins and proteins associated with protein modification, such as prolyl isomerases and proline racemase. The flagellar filament protein, FliC, was down-regulated, possibly as an energy conservation measure, as was the SecA1 preprotein translocase. The up-regulation of hydrogenases and various oxidoreductases suggests that electron flux across these pools of enzymes changes under heat stress. This work represents the first comparative proteomic analysis of the heat stress response in C. difficile strain 630, complementing the existing proteomics data sets and the single microarray comparative analysis of stress response. Thus we have a benchmark proteome for this pathogen, leading to a deeper understanding of its physiology and metabolism informed by the unique functional and adaptive processes used during a temperature upshift mimicking host pyrexia.
Additional Information
© 2011 American Chemical Society. Received: December 9, 2010. Published: July 25, 2011. Published In Issue September 02, 2011. R.L.J.G. was supported by the Northern Ireland Centre of Excellence in Functional Genomics, with funding from the European Union (EU) Programme for Peace and Reconciliation, under the Technology Support for the Knowledge-Based Economy. S.J. was supported by a Vice Chancellor's Research Scholarship award (2007-2010) from the University of Ulster.Attached Files
Supplemental Material - pr200327t_si_001.pdf
Supplemental Material - pr200327t_si_002.xls
Supplemental Material - pr200327t_si_003.pdf
Supplemental Material - pr200327t_si_004.pdf
Supplemental Material - pr200327t_si_005.pdf
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Additional details
- Eprint ID
- 26597
- DOI
- 10.1021/pr200327t
- Resolver ID
- CaltechAUTHORS:20111005-103631419
- Northern Ireland Centre of Excellence in Functional Genomics
- European Union (EU) Programme for Peace and Reconciliation
- University of Ulster
- Created
-
2011-10-05Created from EPrint's datestamp field
- Updated
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2021-11-09Created from EPrint's last_modified field