Correlation of phenotypic profiles using targeted proteomics identifies mycobacterial esx-1 substrates.
Correlation of phenotypic profiles using targeted proteomics identifies mycobacterial esx-1 substrates.
复制标题
DOI：
10.1021/pr500484w
复制
发表时间：
2014-11-07
影响因子：
4.4
通讯作者：
Champion, Patricia A. DiGiuseppe
中科院分区：
文献类型：
--
作者：
Champion, Matthew M.;Williams, Emily A.;Pinapati, Richard S.;Champion, Patricia A. DiGiuseppe
Esx/WXG-100（ESAT-6/Wss）出口商是多蛋白复合物，可促进多种致病性和非致病性细菌物种的蛋白质跨细胞质膜易位。Esx-1（ESAT-6 System-1）系统介导分枝杆菌病原体（包括人类病原体结核分枝杆菌）中的毒力因子易位。虽然已经有几个基因与Esx-1介导的运输和毒力相关，但单个Esx-1基因对输出的贡献在很大程度上是不确定的。Esx-1导出的一个独特之处在于，几种底物需要彼此才能导出/稳定。我们利用底物“相互依赖性”来鉴定Esx-1底物。我们同时定量的13 Esx-1蛋白的水平的变化，从分泌和胞浆蛋白组分产生的16 Esx-1缺陷的海分枝杆菌菌株在一个单一的实验中，使用MRM/SRM靶向质谱。这种可测量的Esx-1蛋白的扩展使我们能够定义使用已知Esx-1底物的表型谱分配新底物的统计规则。使用这种方法，我们确定了三个额外的esx-1基板编码的esx-1区域。我们的研究开始解决特定基因的破坏如何影响Esx-1复合物中的几种蛋白质。总的来说，我们的发现阐明了Esx-1蛋白之间的关系，并为识别适用于其他蛋白质输出体和途径的分泌底物创建了一个框架。
The Esx/WXG-100 (ESAT-6/Wss) exporters are multiprotein complexes that promote protein translocation across the cytoplasmic membrane in a diverse range of pathogenic and nonpathogenic bacterial species. The Esx-1 (ESAT-6 System-1) system mediates virulence factor translocation in mycobacterial pathogens, including the human pathogen Mycobacterium tuberculosis. Although several genes have been associated with Esx-1-mediated transport and virulence, the contribution of individual Esx-1 genes to export is largely undefined. A unique aspect of Esx-1 export is that several substrates require each other for export/stability. We exploited substrate “codependency” to identify Esx-1 substrates. We simultaneously quantified changes in the levels of 13 Esx-1 proteins from both secreted and cytosolic protein fractions generated from 16 Esx-1-deficient Mycobacterium marinum strains in a single experiment using MRM/SRM targeted mass spectrometry. This expansion of measurable Esx-1 proteins allowed us to define statistical rules for assigning novel substrates using phenotypic profiles of known Esx-1 substrates. Using this approach, we identified three additional Esx-1 substrates encoded by the esx-1 region. Our studies begin to address how disruption of specific genes affects several proteins in the Esx-1 complex. Overall, our findings illuminate relationships between Esx-1 proteins and create a framework for the identification of secreted substrates applicable to other protein exporters and pathways.
登录
查看更多内容
影响因子：
3.6
作者：
Coros, Abbie;Callahan, Brian;Derbyshire, Keith M.
通讯作者：
Derbyshire, Keith M.
DOI：
10.1074/mcp.m112.017533
发表时间：
2012-09
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
Champion MM;Williams EA;Kennedy GM;Champion PA
通讯作者：
Champion PA
影响因子：
3.6
作者：
Chen, Jeffrey M.;Zhang, Ming;Cole, Stewart T.
通讯作者：
Cole, Stewart T.
影响因子：
3.2
作者：
Chen, Jeffrey M.;Zhang, Ming;Cole, Stewart T.
通讯作者：
Cole, Stewart T.
DOI：
10.1073/pnas.1119453109
发表时间：
2012-07-10
影响因子：
11.1
作者：
Daleke, Maria H.;Ummels, Roy;Bitter, Wilbert
通讯作者：
Bitter, Wilbert