Proteomic and Transcriptomic Profiling of Staphylococcus aureus Surface LPXTG-proteins: Correlation with agr Genotypes and Adherence Phenotypes

摘要: Staphylococcus aureus infections involve numerous adhesins and toxins, which expression depends on complex regulatory networks. Adhesins include a family of surface proteins covalently attached to the peptidoglycan via conserved LPXTG motif. Here we determined protein mRNA LPXTG-proteins S. Newman in time-course experiments, their relation fibrinogen adherence vitro. Experiments were performed with mutants global accessory-gene regulator (agr), A (Spa), fibrinogen-binding (ClfA), as well during growth iron-rich or iron-poor media. Surface recovered by trypsin-shaving live bacteria. Released peptides analyzed liquid chromatography coupled tandem mass-spectrometry. To unambiguously identify unique LPXTG-proteins, analytical conditions refined using reference library heterogeneously expressed surrogate Lactococcus lactis. Transcriptomes microarrays. Sixteen 18 present detected proteomics. Nine showed bell-shape agr-like that was abrogated agr-negative including Spa, fibronectin-binding (FnBPA), ClfA, iron-binding IsdA, IsdB, immunomodulator SasH, functionally uncharacterized SasD, biofilm-related SasG methicillin resistance-related FmtB. However, only Spa SasH modified proteomic profiles parallel parent its agr- mutant, whereas all other independently mRNA. Moreover, ClfA became highly transcribed active fibrinogen-adherence tests late (24 h), it remained poorly On hand, iron-regulated IsdA-B-C increased >10-times conditions. Thus, proteomic, transcriptomic, adherence-phenotype demonstrated differential aureus. trypsin peptide signatures suggested domain exposures various environments, might be relevant for anti-adhesin vaccines. comprehensive understanding physiology should integrate three approaches.