Proteome analysis of an attenuated Francisella tularensis dsbA mutant: identification of potential DsbA substrate proteins
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
19799467
DOI
10.1021/pr900570b
Knihovny.cz E-resources
- MeSH
- Bacterial Proteins * chemistry genetics metabolism MeSH
- Cell Line MeSH
- Chromatography, Liquid methods MeSH
- Virulence Factors chemistry genetics metabolism MeSH
- Francisella tularensis * genetics metabolism pathogenicity MeSH
- Mass Spectrometry methods MeSH
- Isoelectric Focusing MeSH
- Humans MeSH
- Macrophages cytology metabolism MeSH
- Membrane Proteins * chemistry genetics metabolism MeSH
- Survival Rate MeSH
- Molecular Sequence Data MeSH
- Mutation MeSH
- Mice, Inbred BALB C MeSH
- Mice MeSH
- Peptides chemistry genetics metabolism MeSH
- Protein Disulfide-Isomerases * chemistry genetics metabolism MeSH
- Proteome analysis MeSH
- Proteomics methods MeSH
- Tularemia metabolism mortality MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Bacterial Proteins * MeSH
- Virulence Factors MeSH
- Membrane Proteins * MeSH
- Peptides MeSH
- Protein Disulfide-Isomerases * MeSH
- Proteome MeSH
Francisella tularensis (F. tularensis) is highly infectious for humans via aerosol route and untreated infections with the highly virulent subsp. tularensis can be fatal. Our knowledge regarding key virulence determinants has increased recently but is still somewhat limited. Surface proteins are potential virulence factors and therapeutic targets, and in this study, we decided to target three genes encoding putative membrane lipoproteins in F. tularensis LVS. One of the genes encoded a protein with high homology to the protein family of disulfide oxidoreductases DsbA. The two other genes encoded proteins with homology to the VacJ, a virulence determinant of Shigella flexneri. The gene encoding the DsbA homologue was verified to be required for survival and replication in macrophages and importantly also for in vivo virulence in the mouse infection model for tularemia. Using a combination of classical and shotgun proteome analyses, we were able to identify several proteins that accumulated in fractions enriched for membrane-associated proteins in the dsbA mutant. These proteins are substrate candidates for the DsbA disulfide oxidoreductase as well as being responsible for the virulence attenuation of the dsbA mutant.
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