title: The Enterococcus faecalis exoproteome: identification and temporal regulation by Fsr.
abstract: Analysis of the culture supernatant exoproteins produced by two PFGE clusters of high-level gentamicin and ciprofloxacin-resistant clinical isolates of Enterococcus faecalis from the UK and Ireland revealed two distinct protein profiles. This grouping distinguished OG1RF and GelE metalloprotease-expressing isolates from JH2-2 and other GelE-negative isolates. The integrity of the fsrABDC operon was found to determine the exoproteome composition, since an fsrB mutant of strain OG1RF appeared very similar to that of strain JH2-2, and complementation of the latter with the fsrABDC operon produced an OG1RF-like exoproteome. The proteins present in the supernatant fraction of OG1RF were separated using 2D gels and identified by mass spectrometry and comprised many mass and pI variants of the GelE and SprE proteases. In addition cell wall synthesis and cell division proteins were identified. An OG1RF fsrB mutant had a distinct exoprotein fraction with an absence of the Fsr-regulated proteases and was characterised by general stress and glycolytic proteins. The exoproteome of the OG1RF fsrB mutant resembles that of a divIVA mutant of E. faecalis, suggestive of a stress phenotype.
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Jayendra
authors.initials: J
authors.lastname: Shankar
authors.forename: Rachel G
authors.initials: RG
authors.lastname: Walker
authors.forename: Deborah
authors.initials: D
authors.lastname: Ward
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Horsburgh
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D013293:Enterococcus faecalis / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D009876:Operon / Q000235:genetics; D010449:Peptide Mapping; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; C409031:FsrA protein, Enterococcus faecalis; D020543:Proteome