Protein Kinase Signaling Pathways

蛋白激酶信号通路

基本信息

批准号：
7462243
负责人：
LEWIS C. CANTLEY
金额：
$ 34.25万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7462243
关键词：
Acetylation Amino Acids Arginine Binding Binding Sites C2 Domain Catalytic Domain Cell Survival Complex Cyclin-Dependent Kinases Databases Family member Grant Hydroxylation Individual Internet Knowledge Laboratories Link Location Lysine Mediating Methylation Modification Nuclear Protein Nuclear Proteins Orthologous Gene PTB Domain Pathway interactions Peptide Library Peptides Phosphopeptides Phosphoproteins Phosphorylation Phosphorylation Site Phosphotransferases Polo-Box Domain Post-Translational Protein Processing Proline Protein Binding Domain Protein Databases Protein Kinase Proteins Proteome Research Roentgen Rays Screening procedure Signal Pathway Signal Transduction Signaling Protein Site Specificity Structure Techniques Technology Tertiary Protein Structure base cell growth improved in vivo interest novel programs protein aminoacid sequence protein function

项目摘要

DESCRIPTION (provided by applicant): Intracellular signaling networks are primarily controlled by a complex array of covalent protein modifications that either reversibly or irreversibly alter protein function, multimerization, location, or all three. The most well studied protein modification is phosphorylation, yet our understanding of how specificity is maintained in protein kinase phosphorylation cascades is poorly understood. The hypothesis guiding this study is that domains on signaling proteins mediate assembly into specific complexes via interaction with relatively short sequence motifs on target proteins. Additional fidelity is derived by the specificity of protein kinase catalytic domains for unique substrate motifs. Thus, subtle differences between the motifs recognized by the catalytic pocket of protein kinases and/or differences between the motifs recognized by protein interaction domains outside the catalytic pocket can ultimately explain how individual protein kinases phosphorylate unique sites in vivo. Peptide library screening techniques developed in this laboratory over the past 12 years make it feasible to rapidly deduce the motifs recognized by individual protein interaction domains and motifs phosphorylated by protein kinases. The aims of this grant are: 1) to use a novel peptide library approach to define optimal substrates for protein kinases implicated in cell growth pathways; 2) to use phosphopeptide motif mixtures to identify new phosphoprotein binding domains; 3) to use a similar approach to identify protein domains that bind to other types of modified amino acids; and 4) to further develop our Internet-based Scansite program for predicting sites of protein modification and protein interactions based on sequences in protein databases.

描述（由申请人提供）：细胞内信号网络主要由一系列复杂的共价蛋白质修饰控制，这些修饰可逆或不可逆地改变蛋白质功能、多聚化、位置或全部三者。研究得最多的蛋白质修饰是磷酸化，但我们对蛋白激酶磷酸化级联中如何保持特异性的了解却知之甚少。指导这项研究的假设是信号蛋白上的结构域通过与靶蛋白上相对较短的序列基序相互作用来介导组装成特定的复合物。额外的保真度源自蛋白激酶催化结构域对独特底物基序的特异性。因此，蛋白激酶催化口袋识别的基序之间的细微差异和/或催化口袋外部的蛋白质相互作用结构域识别的基序之间的差异可以最终解释单个蛋白激酶如何磷酸化体内的独特位点。该实验室过去 12 年开发的肽库筛选技术使得快速推断单个蛋白质相互作用结构域识别的基序和蛋白激酶磷酸化的基序成为可能。该资助的目的是：1）使用新型肽库方法来定义细胞生长途径中涉及的蛋白激酶的最佳底物； 2) 使用磷酸肽基序混合物来鉴定新的磷蛋白结合域； 3) 使用类似的方法来鉴定与其他类型的修饰氨基酸结合的蛋白质结构域； 4) 进一步开发基于互联网的 Scansite 程序，用于根据蛋白质数据库中的序列预测蛋白质修饰和蛋白质相互作用的位点。