Dynamics of inhibitor binding and regulation of protein tyrosine kinases

项目摘要。蛋白激酶是人类细胞中普遍存在的信号传导酶的大家族。他们的 失调通常是癌症等疾病的基础，使它们成为极好的治疗靶点，当药物 可以实现特异性。然而，蛋白激酶的高度结构和序列保守性 催化域使特异性抑制剂的开发复杂化。少数临床上成功的激酶 抑制剂部分地通过仅结合不同的激酶构象而实现特异性。虽然分析 数以千计的蛋白激酶的X射线晶体结构表明，单个激酶结构域可以进入 不同的活性和非活性构象，很少有人知道激酶之间如何相互转换， 构象这一建议的理由是，定量了解这些稳定性， 构象及其相互转化的动力学是理解激酶活性、调节和 健康和疾病状态中的配体结合。 本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的激酶结构域内的相互转化和混杂的药物结合 酪氨酸激酶DDR 1。这项建议是围绕四个问题进行的连续研究的一部分， 关注蛋白激酶的构象动力学在激酶调节（Q1）、变构调节 (Q2)配体结合动力学（Q3）和药物特异性/激酶混杂性（Q4）： 什么是酪氨酸激酶构象交换的热力学和动力学？ 问题2：变构信号是如何通过蛋白质结构域传递的，变构信号的结合位点是如何通过蛋白质结构域传递的？ 监管机构预测？ Q3：配体结合动力学的分子决定因素是什么？ Q4：为什么有些激酶会杂乱地结合抑制剂，具有细胞效力的特异性抑制剂是如何结合的？ 发达？ PI和他的团队将通过结构方法（X射线和NMR）的组合来研究这些问题， 配体结合动力学、蛋白质工程、化学生物学和计算方法。网络 富有成效的合作支持了这个项目。该项目的影响是为临床医生提供 耐药突变的机制，细胞生物学家与参数，以了解激酶信号和药物 化学家用参数来调节配体结合动力学。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.