Chemical Approaches to Characterizing Kinase-catalyzed Modifications

描述（由申请人提供）：磷酸化是一种普遍存在的翻译后蛋白质修饰，在多种细胞功能中起着关键作用。事实上，针对磷酸化蛋白底物的激酶的各种药物已经被开发出来，用于治疗从癌症到阿尔茨海默病的疾病。因此，对细胞中激酶及其磷酸化蛋白底物的监测是表征健康和疾病状态以及开发新药的重要目标。不幸的是，现有的方法来监测细胞磷酸化是具有挑战性的。监测和表征激酶及其修饰底物的新方法对于充分理解细胞信号传导和疾病形成是必要的。该项目的长期目标是严格表征激酶活性和激酶催化修饰。该项目的科学基础是我们最近发现的激酶混杂接受？-磷酸修饰ATP类似物作为辅底物。基于激酶ATP共底物的混杂，我们正在追求两个重要的目标。首先，我们假设除了磷酸化之外，激酶还催化蛋白质修饰。我们的假设是基于几个？-磷酸修饰的ATP类似物是自然存在的，与细胞应激条件有关，尽管其作用机制尚不清楚。与我们的假设一致，我们最近发现了一种新的翻译后蛋白质修饰，称为核苷酸磷酸化。由于它们在细胞信号传导中的关键作用，这些研究对于阐明激酶在细胞生物学中的整体作用至关重要。我们的第二个重要目标集中在用各种功能探针标记激酶底物的实验上，以允许检测，序列鉴定和激酶催化修饰的表征。通过使用已开发的化学方法来监测和表征激酶在健康和病变组织中的作用，整个项目的重要成果是对疾病形成中的信号传导有了更深入的了解。因为激酶是药物开发的目标，这些研究将导致创造更有效的影响蛋白质磷酸化的药物。公共卫生相关性：激酶与多种疾病有关，是多种药物的靶标。激酶使蛋白质底物磷酸化，使磷酸化蛋白的表征成为健康相关研究的一个重要方面。该应用概述了表征激酶底物和激酶催化修饰的新方法，以帮助理解疾病和开发新的治疗方法。

DESCRIPTION (provided by applicant): Phosphorylation is a ubiquitous post-translational protein modification that plays a critical role in a variety of cellular functions. In fact, various drugs targeting the kinases that phosphorylate protein substrates have been developed to treat diseases from cancers to Alzheimer's disease. As a result, the monitoring of kinases and their phosphoprotein substrates in cells represents an important goal towards characterizing healthy and disease states and development of new drugs. Unfortunately, available methods to monitor cellular phosphorylation are challenging. New methods for monitoring and charactering kinases and their modified substrates are necessary to allow a full understanding of cellular signaling and disease formation. The long-term goal of this project is a rigorous characterization of kinase activity and kinase-catalyzed modifications. The scientific foundation of the project is our recent revelation that kinases promiscuously accept ?-phosphate modified ATP analogs as cosubstrates. Based on kinase ATP cosubstrate promiscuity, we are pursuing two significant aims. First, we hypothesize that kinases catalyze protein modifications in addition to phosphorylation. Our hypothesis is based on the fact that several ?-phosphate modified ATP analogues are naturally occurring and associated with cell stress conditions, although their mechanisms of action are unknown. Consistent with our hypothesis, we have recently discovered a novel post-translational protein modification, termed nucleotide phosphorylation. With their key role in cell signaling, these studies are critical to illuminate the overall role of kinases in cell biology. Our second significant aim centers on experiments to label kinase substrates with various functional probes to allow detection, sequence identification and characterization of the kinase-catalyzed modifications. By using the developed chemical approaches to monitor and characterize the role of kinases in healthy and diseased tissues, the significant outcome of the overall project is a greater understanding of signaling in disease formation. Because kinase enzymes are targeted in drug development, these studies will lead in the creation of more effective drugs influencing protein phosphorylation. PUBLIC HEALTH RELEVANCE: Kinase enzymes are involved in various diseases and are the target of multiple pharmaceutical drugs. Kinases phosphorylate protein substrates, making a characterization of phosphoproteins an important aspect of health-related research. The application outlines new approaches to characterizing kinase substrates and kinase-catalyzed modifications to aid in the understanding of disease and development of new treatments.