Phosphoproteome analysis of E-coli reveals evolutionary conservation of bacterial Ser/Thr/Tyr phosphorylation
Phosphoproteome analysis of E-coli reveals evolutionary conservation of bacterial Ser/Thr/Tyr phosphorylation
复制标题
DOI：
10.1074/mcp.m700311-mcp200
复制
发表时间：
2008-02-01
影响因子：
7
通讯作者：
Mann, Matthias
中科院分区：
文献类型：
--
作者：
Macek, Boris;Gnad, Florian;Mann, Matthias
丝氨酸、苏氨酸和酪氨酸（Ser/Thr/Tyr）上的蛋白磷酸化通常被认为是真核细胞中主要的调节性翻译后修饰。在基因组和蛋白质组水平上越来越多的证据表明，这种修饰也存在于原核生物中并具有功能。我们最近报道了来自革兰氏阳性菌枯草芽孢杆菌模型的第一个深度磷酸化位点解析数据集，表明Ser/Thr/Tyr磷酸化也存在于许多必需的细菌蛋白上。为了测试这种修饰是否在真细菌中很常见，在这里，我们使用最近开发的基于磷酸肽富集和高精度MS的蛋白质组学方法来分析模型革兰氏阴性菌大肠杆菌的磷酸化蛋白质组。在79个E.大肠杆菌蛋白的Ser/Thr/Tyr磷酸化位点分布为68%/23%/9%。尽管它们的系统发育距离远，但E. coli和B.枯草芽孢杆菌的蛋白质在大小、磷酸化类型和Ser/Thr/Tyr磷酸化位点的分布上具有惊人的相似性。通过结合这两个数据集，我们创建了迄今为止最大的细菌磷蛋白磷酸化位点解析数据库（可在www.phosida.com获得），并使用它来研究细菌磷蛋白和磷酸化位点在系统发育树中的进化保守性。我们证明，细菌磷蛋白和磷酸化残基显着更保守的比他们的nonphosphorylated同行，与一些潜在的磷酸化位点保守从古细菌到人类。我们的研究结果建立了Ser/Thr/Tyr磷酸化作为一种常见的翻译后修饰在真细菌中，本细胞生命的开始。
Protein phosphorylation on serine, threonine, and tyrosine (Ser/Thr/Tyr) is generally considered the major regulatory posttranslational modification in eukaryotic cells. Increasing evidence at the genome and proteome level shows that this modification is also present and functional in prokaryotes. We have recently reported the first in-depth phosphorylation site-resolved dataset from the model Gram-positive bacterium, Bacillus subtilis, showing that Ser/Thr/Tyr phosphorylation is also present on many essential bacterial proteins. To test whether this modification is common in Eubacteria, here we use a recently developed proteomics approach based on phosphopeptide enrichment and high accuracy MS to analyze the phosphoproteome of the model Gram-negative bacterium Escherichia coli. We report 81 phosphorylation sites on 79 E. coli proteins, with distribution of Ser/Thr/Tyr phosphorylation sites 68%/23%/9%. Despite their phylogenetic distance, phosphoproteomes of E. coli and B. subtilis show striking similarity in size, classes of phosphorylated proteins, and distribution of Ser/Thr/Tyr phosphorylation sites. By combining the two datasets, we created the largest phosphorylation site-resolved database of bacterial phosphoproteins to date (available at www.phosida.com) and used it to study evolutionary conservation of bacterial phosphoproteins and phosphorylation sites across the phylogenetic tree. We demonstrate that bacterial phosphoproteins and phosphorylated residues are significantly more conserved than their nonphosphorylated counterparts, with a number of potential phosphorylation sites conserved from Archaea to humans. Our results establish Ser/Thr/Tyr phosphorylation as a common posttranslational modification in Eubacteria, present since the onset of cellular life.