IDENTIFICATION OF KINASE SUBSTRATES BY COVALENT CAPTURE OF PHOSPHOPEPTIDES

通过共价捕获磷酸肽来鉴定激酶底物

• 批准号: 8363793
• 负责人: KEVAN M. SHOKAT
• 金额: $ 1.69万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363793
• 关键词: Engineering; Funding; Grant; Label; Mass Spectrum Analysis; Methodology; Methods; National Center for Research Resources; Pathway interactions; Peptides; Phosphopeptides; Phosphotransferases; Principal Investigator; Protein Kinase; Proteins; Research; Research Infrastructure; Resources; Source; Techniques; United States National Institutes of Health; Work; analog; cost; inorganic phosphate; mTOR Signaling Pathway; rapid technique

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. We will optimize a method for rapid identification of protein kinase substrates. In this method a protein kinase is first engineered to accept an ATP analog that allows it to uniquely label its substrate with a bio-orthogonal phosphate analog. Then we will use a highly specific covalent capture-and-release methodology to rapidly purify tagged peptides derived from labeled substrate proteins. Initial work will focus on the optimization of this technique, followed by application to several kinases in several important pathways including the PI3K mTOR signalling pathway.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我们将优化快速鉴定蛋白激酶底物的方法。 在这种方法中，蛋白激酶首先被设计为接受 ATP 类似物，使其能够用生物正交磷酸盐类似物独特地标记其底物。 然后，我们将使用高度特异性的共价捕获和释放方法来快速纯化来自标记底物蛋白的标记肽。 初步工作将集中于该技术的优化，随后应用于包括 PI3K mTOR 信号通路在内的几个重要通路中的几种激酶。