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Two-component signal transduction system SaeRS positively regulates Staphylococcus epidermidis glucose metabolism.

Lou Q, Qi Y, Ma Y, Qu D - ScientificWorldJournal (2014)

Bottom Line: Of 55 identified proteins that significantly differed in expression between the two strains, 15 were upregulated and 40 were downregulated.The downregulated proteins included enzymes related to glycolysis and TCA cycle, suggesting that glucose is not properly utilized in S. epidermidis when saeRS was deleted.The study will be helpful for treatment of S. epidermidis infection from the viewpoint of metabolic modulation dependent on two-component signal transduction system SaeRS.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Henan University, Kaifeng 475004, China.

ABSTRACT
Staphylococcus epidermidis, which is a causative pathogen of nosocomial infection, expresses its virulent traits such as biofilm and autolysis regulated by two-component signal transduction system SaeRS. In this study, we performed a proteomic analysis of differences in expression between the S. epidermidis 1457 wild-type and saeRS mutant to identify candidates regulated by saeRS using two-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/lonization mass spectrometry (MALDI-TOF-MS). Of 55 identified proteins that significantly differed in expression between the two strains, 15 were upregulated and 40 were downregulated. The downregulated proteins included enzymes related to glycolysis and TCA cycle, suggesting that glucose is not properly utilized in S. epidermidis when saeRS was deleted. The study will be helpful for treatment of S. epidermidis infection from the viewpoint of metabolic modulation dependent on two-component signal transduction system SaeRS.

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Identified spots on the 2D proteome map of SAE and WT. SAE and WT were grown in TSB medium at 37°C until the postexponential growth phase; the bacteria were then separated by centrifugation. Bacteria cell pellets were dissolved in lysis buffer and sonicated on ice. The 2-DE gels were performed using 24 cm immobilized dry strips (IPG, nonlinear, pH 4–7, GE Healthcare) and analyzed by ImageMaster 2D platinum 6.0 software (Amersham Biosciences). Differentially expressed protein spots on 2-DE gels were identified by MS/MS analysis and shown in gels with unique protein spot numbers. Protein spots were identified using a 4700 MALDI-TOF/TOF Proteomics Analyzer (Applied Biosystems, California, USA).
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fig1: Identified spots on the 2D proteome map of SAE and WT. SAE and WT were grown in TSB medium at 37°C until the postexponential growth phase; the bacteria were then separated by centrifugation. Bacteria cell pellets were dissolved in lysis buffer and sonicated on ice. The 2-DE gels were performed using 24 cm immobilized dry strips (IPG, nonlinear, pH 4–7, GE Healthcare) and analyzed by ImageMaster 2D platinum 6.0 software (Amersham Biosciences). Differentially expressed protein spots on 2-DE gels were identified by MS/MS analysis and shown in gels with unique protein spot numbers. Protein spots were identified using a 4700 MALDI-TOF/TOF Proteomics Analyzer (Applied Biosystems, California, USA).

Mentions: Two-dimensional gel electrophoresis (2-DE) was performed to analyze the expression of S. epidermidis proteins regulated by two-component signal transduction system SaeRS. The gel images were compared and analyzed. The patterns of protein expression in SAE and WT appeared largely similar but some clear differences in protein expression levels of certain protein spots were evident (Figure 1). Fifty-five protein spots were excised for further analysis by the LC-MS/MS. Of these, 15 spots were up-regulated, and 40 were downregulated (Table 2).


Two-component signal transduction system SaeRS positively regulates Staphylococcus epidermidis glucose metabolism.

Lou Q, Qi Y, Ma Y, Qu D - ScientificWorldJournal (2014)

Identified spots on the 2D proteome map of SAE and WT. SAE and WT were grown in TSB medium at 37°C until the postexponential growth phase; the bacteria were then separated by centrifugation. Bacteria cell pellets were dissolved in lysis buffer and sonicated on ice. The 2-DE gels were performed using 24 cm immobilized dry strips (IPG, nonlinear, pH 4–7, GE Healthcare) and analyzed by ImageMaster 2D platinum 6.0 software (Amersham Biosciences). Differentially expressed protein spots on 2-DE gels were identified by MS/MS analysis and shown in gels with unique protein spot numbers. Protein spots were identified using a 4700 MALDI-TOF/TOF Proteomics Analyzer (Applied Biosystems, California, USA).
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3921950&req=5

fig1: Identified spots on the 2D proteome map of SAE and WT. SAE and WT were grown in TSB medium at 37°C until the postexponential growth phase; the bacteria were then separated by centrifugation. Bacteria cell pellets were dissolved in lysis buffer and sonicated on ice. The 2-DE gels were performed using 24 cm immobilized dry strips (IPG, nonlinear, pH 4–7, GE Healthcare) and analyzed by ImageMaster 2D platinum 6.0 software (Amersham Biosciences). Differentially expressed protein spots on 2-DE gels were identified by MS/MS analysis and shown in gels with unique protein spot numbers. Protein spots were identified using a 4700 MALDI-TOF/TOF Proteomics Analyzer (Applied Biosystems, California, USA).
Mentions: Two-dimensional gel electrophoresis (2-DE) was performed to analyze the expression of S. epidermidis proteins regulated by two-component signal transduction system SaeRS. The gel images were compared and analyzed. The patterns of protein expression in SAE and WT appeared largely similar but some clear differences in protein expression levels of certain protein spots were evident (Figure 1). Fifty-five protein spots were excised for further analysis by the LC-MS/MS. Of these, 15 spots were up-regulated, and 40 were downregulated (Table 2).

Bottom Line: Of 55 identified proteins that significantly differed in expression between the two strains, 15 were upregulated and 40 were downregulated.The downregulated proteins included enzymes related to glycolysis and TCA cycle, suggesting that glucose is not properly utilized in S. epidermidis when saeRS was deleted.The study will be helpful for treatment of S. epidermidis infection from the viewpoint of metabolic modulation dependent on two-component signal transduction system SaeRS.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular and Molecular Immunology, Henan University, Kaifeng 475004, China.

ABSTRACT
Staphylococcus epidermidis, which is a causative pathogen of nosocomial infection, expresses its virulent traits such as biofilm and autolysis regulated by two-component signal transduction system SaeRS. In this study, we performed a proteomic analysis of differences in expression between the S. epidermidis 1457 wild-type and saeRS mutant to identify candidates regulated by saeRS using two-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/lonization mass spectrometry (MALDI-TOF-MS). Of 55 identified proteins that significantly differed in expression between the two strains, 15 were upregulated and 40 were downregulated. The downregulated proteins included enzymes related to glycolysis and TCA cycle, suggesting that glucose is not properly utilized in S. epidermidis when saeRS was deleted. The study will be helpful for treatment of S. epidermidis infection from the viewpoint of metabolic modulation dependent on two-component signal transduction system SaeRS.

Show MeSH
Related in: MedlinePlus