Phosphoproteome analysis of the pathogenic bacterium Helicobacter pylori reveals over-representation of tyrosine phosphorylation and multiply phosphorylated proteins
Phosphoproteome analysis of the pathogenic bacterium Helicobacter pylori reveals over-representation of tyrosine phosphorylation and multiply phosphorylated proteins
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致病菌幽门螺杆菌的磷酸化蛋白质组分析揭示了酪氨酸磷酸化和多重磷酸化蛋白质的过度表达
DOI：
10.1002/pmic.201000649
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发表时间：
2011-04-01
期刊：
影响因子：
3.4
通讯作者：
He, Qing-Yu
中科院分区：
文献类型：
--
作者：
Ge, Ruiguang;Sun, Xuesong;He, Qing-Yu
越来越多的证据表明，丝氨酸(Ser)、苏氨酸(Thr)和酪氨酸(Tyr)残基上的蛋白质磷酸化是细菌中一种主要的调节翻译后修饰。为了揭示革兰氏阴性致病菌幽门螺杆菌的磷酸化状态，我们进行了基于二氧化钛-磷酸肽富集法和高精度LC-MS/MS测定的全局和定点磷酸蛋白质组分析。共鉴定了67种蛋白质的82个磷酸化多肽，共有126个磷酸化位点，其中79个I类位点分别具有42.8：38.7：18.5%的Ser/Thr/Tyr磷酸化。幽门螺杆菌的磷酸化蛋白质组具有相对较大的大小、较高的Tyr磷酸化率、单个磷酸肽中多个磷酸化位点的高丰度以及膜蛋白的过度表达。构建了一个覆盖28个磷酸蛋白的相互作用网络，共163个蛋白，以幽门螺杆菌的主要毒力因子VacA为核心，表明幽门螺杆菌的蛋白磷酸化可能被精细地调控，以调节代谢途径和细菌毒力的许多方面。
Increasing evidence shows that protein phosphorylation on serine (Ser), threonine (Thr) and tyrosine (Tyr) residues is a major regulatory post-translational modification in the bacteria. To reveal the phosphorylation state in the Gram-negative pathogenic bacterium Helicobacter pylori, we carried out a global and site-specific phosphoproteomic analysis based on TiO(2)-phosphopeptide enrichment and high-accuracy LC-MS/MS determination. Eighty-two phosphopeptides from 67 proteins were identified with 126 phosphorylation sites, among which 79 class I sites were determined to have a distribution of 42.8:38.7:18.5% for the Ser/Thr/Tyr phosphorylation, respectively. The H. pylori phosphoproteome is characterized by comparably big size, high ratio of Tyr phosphorylation, high abundance of multiple phosphorylation sites in individual phosphopeptides and over-representation of membrane proteins. An interaction network covering 28 phosphoproteins was constructed with a total of 163 proteins centering on the major H. pylori virulence factor VacA, indicating that protein phosphorylation in H. pylori may be delicately controlled to regulate many aspects of the metabolic pathways and bacterial virulence.