Functional proteomics mapping of a human signaling pathway
Functional proteomics mapping of a human signaling pathway
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DOI：
10.1101/gr.2334104
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发表时间：
2004-07-01
期刊：
影响因子：
7
通讯作者：
Gauthier, JM
中科院分区：
文献类型：
--
作者：
Colland, F;Jacq, X;Gauthier, JM
人类基因组的获取促进了广泛的功能蛋白质组学研究。在这里，我们提出了一种综合的方法，结合大规模的蛋白质相互作用图谱，相互作用网络的探索，以及对新发现的与人类信号通路有关的蛋白质进行的细胞功能分析。作为原理的证明，我们研究了Smad信号系统，它受转化生长因子β(TGFbeta)超家族成员的调控。我们使用双杂交筛选来绘制Smad信号蛋白-蛋白质相互作用的图谱，并建立了一个包含755个相互作用的网络，涉及591个蛋白质，其中179个蛋白质的注释很差或没有注释。使用PIMRider改进了对这种复杂交互数据库的探索，PIMRider是一个可通过Web访问的专用导航工具。18个已知的Smad相关蛋白的存在说明了这个网络的生物学意义。在哺乳动物细胞中进行的功能分析包括siRNA敲除实验确定了八种参与Smad信号转导的新蛋白质，从而验证了这种整合的功能蛋白质组学方法。
Access to the human genome facilitates extensive functional proteomics studies. Here, we present an integrated approach combining large-scale protein interaction mapping, exploration of the interaction network, and cellular functional assays performed on newly identified proteins involved in a human signaling pathway. As a proof of principle, we studied the Smad signaling system, which is regulated by members of the transforming growth factor beta (TGFbeta) superfamily. We used two-hybrid screening to map Smad signaling protein-protein interactions and to establish a network of 755 interactions, involving 591 proteins, 179 of which were poorly or not annotated. The exploration of such complex interaction databases is improved by the use of PIMRider, a dedicated navigation tool accessible through the Web. The biological meaning of this network is illustrated by the presence of 18 known Smad-associated proteins. Functional assays performed in mammalian cells including siRNA knock-down experiments identified eight novel proteins involved in Smad signaling, thus validating this integrated functional proteomics approach.