Dynamics of inhibitor binding and regulation of protein tyrosine kinases

抑制剂结合动力学和蛋白酪氨酸激酶调节

• 批准号: 9313905
• 负责人: Markus A Seeliger
• 金额: $ 39.48万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9313905
• 关键词: Affect; Allosteric Site; Binding; Binding Sites; Biology; Catalytic Domain; Cells; Chemicals; Clinical; Collaborations; Computing Methodologies; Crystallization; Cyclic AMP-Dependent Protein Kinases; DDR1 gene; Development; Disease; Enzymes; Equilibrium; Family; Foundations; Goals; Health; Human; Kinetics; Knowledge; Ligand Binding; Malignant Neoplasms; Methods; Molecular; Molecular Conformation; Pharmaceutical Preparations; Phosphotransferases; Protein Dynamics; Protein Engineering; Protein Kinase; Protein Tyrosine Kinase; Public Health; Regulation; Research; Roentgen Rays; Role; Signal Pathway; Signal Transduction; Specificity; Tertiary Protein Structure; Therapeutic; Thermodynamics; Tyrosine Kinase Domain; design; inhibitor/antagonist; kinase inhibitor; protein structure; resistance mechanism; resistance mutation; src-Family Kinases; therapeutic target

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.

项目摘要。蛋白激酶是人类细胞中普遍存在的信号酶大家族。他们的 失调通常是癌症等疾病的基础，因此当药物 可以实现特异性。然而，蛋白激酶的高度结构和序列保守性 催化结构域使特定抑制剂的开发变得复杂。少数临床上成功的激酶 抑制剂部分通过仅结合不同的激酶构象来实现特异性。虽然分析 数千个蛋白激酶的 X 射线晶体结构表明，单个激酶结构域可以访问 由于不同的活性和非活性构象，人们对激酶如何在不同的活性和非活性构象之间相互转换知之甚少。 构象。该提案的基本原理是对这些稳定性的定量理解 构象及其相互转化的动态是了解激酶活性、调节和作用的关键 健康和疾病状态下的配体结合。 该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换以及混杂的药物结合 酪氨酸激酶 DDR1。该提案是一系列研究的一部分，该研究围绕四个问题： 关注蛋白激酶构象动力学在激酶调节（Q1）、变构调节中的作用 (Q2)、配体结合动力学 (Q3) 和药物特异性/激酶混杂性 (Q4)： Q1：酪氨酸激酶构象交换的热力学和动力学是什么？ 问题 2：变构信号如何通过蛋白质结构域传递以及变构结合位点如何传递 监管者如何预测？ Q3：配体结合动力学的分子决定因素是什么？ Q4：为什么有些激酶会与抑制剂混杂结合，具有细胞效力的特定抑制剂如何结合？ 发达？ PI 和他的团队将通过结构方法（X 射线和核磁共振）的结合来研究这些问题， 配体结合动力学、蛋白质工程、化学生物学和计算方法。一个网络 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 抗性突变机制，细胞生物学家利用参数来了解激酶信号传导和药物 化学家使用参数来调节配体结合动力学。长远目标是为未来打下基础 设计安全有效、足够特异性的疾病相关蛋白激酶抑制剂。