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首页> 外文期刊>BMC Microbiology >Mass spectrometry-based quantitative proteomic analysis of Salmonella enterica serovar Enteritidis protein expression upon exposure to hydrogen peroxide
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Mass spectrometry-based quantitative proteomic analysis of Salmonella enterica serovar Enteritidis protein expression upon exposure to hydrogen peroxide

机译：基于质谱的蛋白质组学蛋白质组学分析肠炎沙门氏菌血清肠炎沙门氏菌暴露于过氧化氢

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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 (H2O2), 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 H2O2 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 H2O2. SPI-1 effector SipC was expressed about 3-fold higher and SopB was expressed approximately 2-fold lower in the presence of H2O2, 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 H2O2. 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.
机译：背景肠道沙门氏菌是一种常见的食源性细菌病原体，据信在存在不同环境压力（例如由过氧化氢（H 2 O 2 ），它是吞噬细胞在感染过程中产生的，代表了宿主细胞的重要抗菌机制。在沙门氏菌蛋白中，沙门氏菌致病岛1和2（SPI-1和SPI-2）的效应子特别受关注，因为它们在体内宿主感染期间表达，并且对于全身性上皮细胞的侵袭和器官复制非常重要。分别感染。然而，尚未广泛研究在确立的系统感染阶段体内这些蛋白质在体内暴露于H 2 O 2 或宿主细胞后的表达情况。结果使用稳定的同位素标记结合质谱，对肠炎沙门氏菌肠炎沙门氏菌进行了蛋白质组学定量分析，鉴定出76种蛋白质，其表达受H 2 O 2 调控。在H 2 O 2 存在下，SPI-1效应子SipC的表达约高3倍，SopB的表达约低2倍，而sipC的表达无明显变化。观察到另一种SPI-1蛋白SipA的表达。 Western分析证实了SipA，SipC和SopB的相对丰度，验证了我们定量分析蛋白质表达方法的准确性和可重复性。此外，免疫检测显示在巨噬细胞和被感染小鼠脾脏的感染后期，SipA和SipC大量表达，但未显示SopB。结论我们已经鉴定了沙门氏菌蛋白，其表达在H 2 O 2 的存在下被调节。我们的研究结果还提供了第一个直接证据，表明SipC在体内沙门氏菌病的晚期在脾脏中高度表达。这些结果表明，SipC和其他调节蛋白在支持沙门氏菌在氧化应激下以及体内全身感染过程中的存活和复制中可能发挥作用。

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