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Subtyping of Campylobacter jejuni ssp. doylei Isolates Using Mass Spectrometry-based PhyloProteomics (MSPP)
Published on: October 30, 2016
Identification of membrane-associated proteins from Campylobacter jejuni strains using complementary proteomics
Stuart J Cordwell1, Alice C L Len, Rachel G Touma
1School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, New South Wales, NSW 2006, Australia. s.cordwell@mmb.usyd.edu.au
Proteomics
|December 21, 2007
Summary
This study comprehensively analyzed Campylobacter jejuni membrane proteins, identifying 453 unique proteins, including 187 bona fide membrane proteins. Key virulence factors like chemotaxis proteins and outer membrane proteins were highlighted for potential roles in gastrointestinal infections.
Area of Science:
- Microbiology
- Proteomics
- Infectious Diseases
Background:
- Campylobacter jejuni is a primary cause of foodborne and waterborne illnesses globally.
- Understanding its membrane proteome is crucial for identifying virulence factors and developing control strategies.
Purpose of the Study:
- To comprehensively analyze the membrane-associated proteome of a virulent Campylobacter jejuni gastrointestinal isolate (JHH1).
- To compare membrane proteins between virulent (JHH1) and less virulent (ATCC 700297) strains to identify potential virulence determinants.
- To identify immunoreactive membrane proteins recognized by patient sera.
Main Methods:
- Sodium carbonate precipitation and ultracentrifugation to isolate membrane proteins.
- Two-dimensional gel electrophoresis (2-DE) and MALDI-TOF MS for initial protein identification.
- Two-dimensional liquid chromatography (2-DLC) coupled with MS/MS for in-depth proteomic analysis.
- Comparative analysis and immunoblotting with patient sera.
Main Results:
- Identification of 453 unique proteins, with 187 confirmed membrane proteins, representing 27.4% of the C. jejuni proteome.
- Discovery of 28 proteins significantly more abundant or unique to the virulent JHH1 strain, including chemotaxis proteins, CjaA, CjaC, and four uncharacterized outer membrane proteins (OMPs).
- Identification of 13 immunoreactive proteins in JHH1 and 12 in ATCC 700297, including flagellin (FlaA), CadF, and several outer membrane proteins.
Conclusions:
- This study provides the most comprehensive membrane proteome analysis of C. jejuni to date.
- Identified virulence-associated proteins, including novel OMPs and chemotaxis-related proteins, offer targets for future research into C. jejuni pathogenesis.
- The findings contribute to a better understanding of C. jejuni infection mechanisms and host immune responses.
