Identification of S-nitrosylation of proteins of Helicobacter pylori in response to nitric oxide stress
Identification of S-nitrosylation of proteins of Helicobacter pylori in response to nitric oxide stress
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DOI：
10.1007/s12275-011-0262-7
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发表时间：
2011-05
期刊：
影响因子：
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通讯作者：
W. Qu;Ya-bin Zhou;Yundong Sun;M. Fang;Han Yu;Wenjuan Li;Zhifang Liu;Jiping Zeng;Chunyan Chen;Chengjiang Gao;J. Jia
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文献类型：
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作者：
W. Qu;Ya-bin Zhou;Yundong Sun;M. Fang;Han Yu;Wenjuan Li;Zhifang Liu;Jiping Zeng;Chunyan Chen;Chengjiang Gao;J. Jia
当幽门螺杆菌侵入胃粘膜时，人类的先天性和适应性免疫应答被激活。一氧化氮（NO）和活性氮是重要的免疫效应物质，通过蛋白质的氧化和S-亚硝基化作用发挥其功能。以S-亚硝基谷胱甘肽和硝普钠为NO供体，H。pylori细胞与这些化合物一起孵育以分析NO的抑制作用。pylori已经在体外显示。此外，在H.幽门螺杆菌的鉴定是通过生物素开关法结合基质辅助激光解吸电离/飞行时间串联质谱（MALDI-TOF-MS/MS）进行的。五个S-亚硝基化的蛋白质鉴定的伴侣和热休克蛋白（GroEL），烷基氢过氧化物还原酶（TsaA），尿素酶α亚基（UreA），HP 0721和HP 0129。重要的是，使用纯化的重组蛋白证实了TsaA和UreA的S-亚硝基化。考虑到这些酶在抗氧化防御、粘附和定殖中的重要性，NO可能通过S-亚硝基化靶向酶发挥其抗菌作用。蛋白质S-亚硝基化的鉴定可能有助于了解NO的抗菌作用。我们的研究结果为开发新的抗H.幽门感染
Innate and adaptive immune responses are activated in humans when Helicobacter pylori invades the gastric mucosa. Nitric oxide (NO) and reactive nitrogen species are important immune effectors, which can exert their functions through oxidation and S-nitrosylation of proteins. S-nitrosoglutathione and sodium nitroprus-side were used as NO donors and H. pylori cells were incubated with these compounds to analyze the inhibitory effect of NO. The suppressing effect of NO on H. pylori has been shown in vitro. Furthermore, the proteins modified by S-nitrosylation in H. pylori were identified through the biotin switch method in association with matrix-assisted laser desorption ionization/time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS). Five S-nitrosylated proteins identified were a chaperone and heat-shock protein (GroEL), alkyl hydroperoxide reductase (TsaA), urease alpha subunit (UreA), HP0721, and HP0129. Importantly, S-nitrosylation of TsaA and UreA were confirmed using purified recombinant proteins. Considering the importance of these enzymes in antioxidant defenses, adherence, and colonization, NO may exert its antibacterial actions by targeting enzymes through S-nitrosylation. Identification of protein S-nitrosylation may contribute to an understanding of the antibacterial actions of NO. Our findings provide an insight into potential targets for the development of novel therapeutic agents against H. pylori infection.