Modeling human phosphorylation networks through kinome-wide profiling

描述（由申请人提供）：蛋白质磷酸化是真核生物中最常见的可逆翻译后修饰，但包含蛋白激酶、其调节剂和其底物的信号传导网络仅部分阐明。拟议项目的总体目标是使用阵列位置扫描肽库为人类基因组中编码的蛋白激酶的整个集合建立一个全面的共识磷酸化基序集合，并将这些数据整合到目前整个生物医学界可用的网络访问工具中。（1）通过现在鉴定负责这些修饰的相关激酶和信号通路，允许对大量蛋白质进行功能注释，所述蛋白质的磷酸化位点已经或目前正在高通量磷酸蛋白质组质谱实验和数据集中被定位，所述数据集先前由NIH资助;（2）允许鉴定新蛋白质 与人类健康和疾病相关的激酶底物，并将它们置于特定信号转导途径的背景下;和（3）提供了一套通用的激酶工具，可用于结构和药物抑制剂研究，并用于治疗靶向与人类疾病有关的特定信号转导途径。为了说明我们的方法的实用性，我们将研究预测的底物蛋白激酶在河马信号通路，一个保守的肿瘤抑制途径，重要的调节细胞增殖，分化和生存。该项目将有助于增加我们对蛋白激酶如何实现特异性的基本理解，将确定信号网络中的关键连接，并将为研究信号转导和蛋白磷酸化的研究人员提供一般资源。

DESCRIPTION (provided by applicant): Protein phosphorylation is the most common reversible post-translational modification in eukaryotes, yet signaling networks comprising protein kinases, their regulators, and their substrates are only partially elucidated. The overall goals of the proposed project are to build a comprehensive collection of consensus phosphorylation motifs for the entire collection of protein kinases encoded in the human genome using arrayed positional scanning peptide libraries, and integrate this data into web-accessible tools that are currently available to the entire biomedical community. The resulting dataset of protein kinase specificity motifs and informatics tools will: (1) allow functional annotation of a large number of proteins whose phosphorylation sites already have been, or currently are, being mapped in high-throughput phosphoproteomic mass- spectrometry experiments and datasets that have been previously funded by the NIH by now identifying the relevant kinase and signaling pathways responsible for these modifications; (2) allow the identification of new protein kinase substrates relevant to human health and disease and place them within the context of specific signal transduction pathways; and (3) provide a general set of kinase tools useful for structural and drug inhibitor studies and for the therapeutic targeting of specific signaling pathways implicated in human disease. To illustrate the utility of our approach, we will investigate predicted substrates of protein kinases in the Hippo signaling pathway, a conserved tumor suppressor pathway important in regulating cell proliferation, differentiation and survival. This project will serve to increase our fundamental understanding of how specificity is achieved by protein kinases, will identify critical connections in signaling networks, and will provide a general resource for researchers studying signal transduction and protein phosphorylation.