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Background   Salmonella   enterica , a common food-borne bacterial pathogen, is believed to change its protein expression profile in the presence of different environmental stress such as that caused by the exposure to hydrogen peroxide (H 2 O 2 ), which can be generated by phagocytes during infection and represents an important antibacterial mechanism of host cells. Among  Salmonella  proteins, the effectors of  Salmonella  pathogenicity island 1 and 2 (SPI-1 and SPI-2) are of particular interest since they are expressed during host infection  in vivo  and are important for invasion of epithelial cells and for replication in organs during systemic infection, respectively. However, the expression profiles of these proteins upon exposure to H 2 O 2  or to host cells  in vivo  during the established phase of systemic infection have not been extensively studied.    Results  Using stable isotope labeling coupled with mass spectrometry, we performed quantitative proteomic analysis of  Salmonella   enterica  serovar Enteritidis and identified 76 proteins whose expression is modulated upon exposure to H 2 O 2 . SPI-1 effector SipC was expressed about 3-fold higher and SopB was expressed approximately 2-fold lower in the presence of H 2 O 2 , while no significant change in the expression of another SPI-1 protein SipA was observed. The relative abundance of SipA, SipC, and SopB was confirmed by Western analyses, validating the accuracy and reproducibility of our approach for quantitative analysis of protein expression. Furthermore, immuno-detection showed substantial expression of SipA and SipC but not SopB in the late phase of infection in macrophages and in the spleen of infected mice.    Conclusions  We have identified  Salmonella  proteins whose expression is modulated in the presence of H 2 O 2 . Our results also provide the first direct evidence that SipC is highly expressed in the spleen at late stage of salmonellosis  in vivo . These results suggest a possible role of SipC and other regulated proteins in supporting survival and replication of  Salmonella  under oxidative stress and during its systemic infection  in vivo .

bmc microbiology

Mass spectrometry-based quantitative proteomic analysis of Salmonella enterica serovar Enteritidis protein expression upon exposure to hydrogen peroxide