IDENTIFICATION OF KINASE SUBSTRATES BY COVALENT CAPTURE OF PHOSPHOPEPTIDES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们将优化一种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成接受一种ATP类似物，从而允许它用生物正交磷酸类似物唯一地标记其底物。然后，我们将使用一种高度特异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多肽。最初的工作将集中在这项技术的优化上，随后将应用于几种具有已知线粒体靶点的激酶。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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 with known mitochondrial targets.