Dynamics of inhibitor binding and regulation of protein tyrosine kinases

项目总结。蛋白激酶是人类细胞中普遍存在的一大类信号转导酶。他们的 调节失调往往是癌症等疾病的基础，使它们成为极好的治疗靶点，当药物 可以实现专一性。然而，蛋白激酶的高度结构和序列保守性 催化结构域使特定抑制剂的开发变得复杂。为数不多的临床成功的激酶 抑制剂通过仅与不同的激酶构象结合来部分实现特异性。在分析的同时 数以千计的蛋白激酶的X射线晶体结构表明，单一的激酶域可以访问 不同的活跃和不活跃的构象，鲜为人知的是， 构象。这一提议的基本原理是，对这些因素的稳定性的量化理解 构象及其相互转化的动力学是理解激酶活性、调控和 在健康和疾病状态下的配基结合。 这个项目的目标是描述构象的动力学和平衡参数 酪氨酸激酶Src、Abl、Brk激活域的相互转换与药物结合的混杂 酪氨酸激酶DDR1。这项建议是围绕四个问题进行的连续研究的一部分，这些问题 关注蛋白激酶构象动力学在蛋白激酶调节(Q1)、变构调节中的作用 (Q2)、配体结合动力学(Q3)和药物特异性/激酶混杂(Q4)： 问1：酪氨酸酶的构象交换的热力学和动力学是什么？ 问题2：变构信号是如何通过蛋白质结构域传递的，以及变构的结合部位是如何传递的 监管机构被预测到了吗？ 问3：配体结合动力学的分子决定因素是什么？ 问4：为什么一些蛋白激酶会混杂地与抑制剂结合？具有细胞效力的特定抑制剂是如何结合的？ 发达吗？ PI和他的团队将通过结构方法(X射线和核磁共振)的组合来研究这些问题， 配基结合动力学、蛋白质工程、化学生物学和计算方法。一个网络 富有成效的协作为该项目提供了支持。该项目的影响是为临床医生提供 耐药突变的机制，细胞生物学家用参数了解激酶信号转导和药物 用参数来调节配基结合动力学的化学家。我们的长期目标是为 设计安全、有效、足够特异的疾病相关蛋白激酶抑制剂。

PROJECT SUMMARY. Protein kinases are a large family of ubiquitous signaling enzymes in human cells. Their dysregulation often underlies diseases such as cancer, making them excellent therapeutic targets, when drug specificity can be achieved. However, the high structural and sequence conservation of the protein kinase catalytic domains has complicated the development of specific inhibitors. The few clinically-successful kinase inhibitors achieve specificity in part by binding only to distinct kinase conformations. While the analysis of thousands of X-ray crystal structures of protein kinases has shown that a single kinase domain can access different active and inactive conformations, little is known about how kinases interconvert between the conformations. The rationale of this proposal is that a quantitative understanding of the stability of these conformations and the dynamics of their interconversion are key to understanding kinase activity, regulation and ligand binding in health and disease states. The objective of this project is to describe the kinetic and equilibrium parameters for the conformational interconversions within the kinase domains of tyrosine kinases Src, Abl, Brk and the promiscuous drug-binding tyrosine kinase DDR1. This proposal is part of a continuum of research centered around four questions that concern the role of conformational dynamics of protein kinases in kinase regulation (Q1), allosteric modulation (Q2), ligand binding kinetics (Q3) and drug specificity/kinase promiscuity (Q4): Q1: What are the thermodynamics and kinetics of conformational exchange in tyrosine kinases? Q2: How are allosteric signals communicated through protein domains and how can binding sites for allosteric regulators be predicted? Q3: What are the molecular determinants of ligand-binding kinetics? Q4: Why do some kinases bind inhibitors promiscuously and how can specific inhibitors with cellular potency be developed? The PI and his team will study these questions through a combination of structural methods (X-ray and NMR), ligand binding kinetics, protein engineering, chemical biology and computational methods. A network of productive collaborations supports this project. The impact of this project is to provide clinicians with the mechanism of resistance mutations, cell biologists with parameters to understand kinase signaling and medicinal chemists with parameters to modulate ligand binding kinetics. The long-term goal is to lay the foundation for the design of safe and effective, sufficiently specific, inhibitors of disease-associated protein kinases.