title: Proteomic analysis of cell surface proteins from Clostridium difficile.
abstract: Clostridium difficile is a bacterium that causes disease of the large intestine, particularly after treatment with antibiotics. The bacterium produces two toxins (A and B) that are responsible for the pathology of the disease. In addition, a number of bacterial virulence factors associated with adhesion to the gut have previously been identified, including the cell wall protein Cwp66, the high-molecular weight surface layer protein (HMW-SLP) and the flagella. As the genome sequence predicts many other cell wall associated proteins, we have investigated the diversity of proteins in cell wall extracts, with the aim of identifying further virulence factors. We have used a number of methods to remove the proteins associated with the cell wall of C. difficile. Two of the resulting extracts, obtained using low pH glycine treatment and lysozyme digestion of the cell wall, have been analysed in detail by two-dimensional electrophoresis and mass spectrometry. One hundred and nineteen spots, comprising 49 different proteins, have been identified. The two surface layer proteins (SLPs) are the most abundant proteins, and we have also found components of the flagellum. Interestingly, we have also determined that a number of paralogs of the HMW-SLP are expressed, and these could represent targets for further investigation as virulence factors.
authors.affiliation: Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College London, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Wright
authors.forename: Robin
authors.initials: R
authors.lastname: Wait
authors.forename: Shajna
authors.initials: S
authors.lastname: Begum
authors.forename: Ben
authors.initials: B
authors.lastname: Crossett
authors.forename: Judit
authors.initials: J
authors.lastname: Nagy
authors.forename: Katherine
authors.initials: K
authors.lastname: Brown
authors.forename: Neil
authors.initials: N
authors.lastname: Fairweather
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D016360:Clostridioides difficile / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005407:Flagella / Q000378:metabolism; D013058:Mass Spectrometry; D020543:Proteome / Q000378:metabolism*; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D020543:Proteome; D037521:Virulence Factors