Decoding the dynamic mechanism of allosteric activation in the cyclin-dependent kinase Cdk2

解读细胞周期蛋白依赖性激酶 Cdk2 变构激活的动态机制

基本信息

批准号：
10321568
负责人：
Nicholas Mark Levinson
金额：
$ 30.8万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10321568
关键词：
Active Sites Address Antineoplastic Agents Binding Biology Case Study Cell Cycle Cell Cycle Progression Cell Proliferation Cell physiology Chemicals Clinic Clinical Complex Coupling Crystallization Cyclic AMP-Dependent Protein Kinases Cyclin-Dependent Kinases Cyclins Data Development Disease Drug Design Elements Equilibrium Eukaryotic Cell Family Fluorescence Resonance Energy Transfer Foundations G1 Phase Goals Human Kinetics Ligands Link Malignant Neoplasms Maps Measures Mediating Methods Molecular Molecular Conformation Motion Nature Nuclear Magnetic Resonance Oncogenic Optics Pharmaceutical Preparations Phase Transition Phosphorylation Phosphotransferases Population Positioning Attribute Process Protein Dynamics Protein Kinase Proteins Regulation Relaxation Resistance Resolution Roentgen Rays Role Sampling Signal Transduction Signaling Protein Spectrum Analysis Structure Testing Therapeutic Work cancer therapy cell growth combat conformational conversion design experimental study inhibitor insight kinase inhibitor monomer next generation novel overexpression public health relevance small molecule targeted treatment time use tumorigenesis

项目摘要

Project Abstract This project reaches towards a new paradigm for how conformational dynamics of protein kinases drive their catalytic activation, and provides an important case study of how protein dynamics can be commandeered to artificially control kinase activity with allosteric small molecules. The protein kinases are a large family of signaling proteins that control cell growth and proliferation in all eukaryotic cells. Kinases behave like molecular switches, transitioning between catalytically active “on” and inactive “off” states in a tightly controlled fashion to bring about prescribed changes in cell physiology. This stringent regulatory control is widely disrupted in cancer, and targeting aberrant kinase activity with small- molecule drugs is now an important component of many cancer treatments. Crystal structures have given us static pictures of the on and off states of kinases, but they tell us little about how the proteins transition between these states, which is an inherently dynamic process. The nature of these dynamic transitions and how they are perturbed in disease remain largely obscure, a fact that has impeded our ability to design allosteric therapeutics that switch kinases to the off state by mimicking their natural control mechanisms. Instead, existing kinase inhibitors work by binding to the highly conserved active site, a mode of action that makes them poorly selective. The goal of this project is to use a combination of experimental methods that provide complementary information about protein dynamics to determine how kinases transition between different conformational states, and to understand how kinase dynamics can be commandeered by small-molecule drugs to artificially modulate kinase function. Using nuclear magnetic resonance and optical spectroscopy applied to the cyclin- dependent kinase Cdk2, a key regulator of cell cycle progression, we aim to reveal 1) the allosteric coupling mechanism that links cyclin binding to activation of Cdk2, 2) how phosphorylation cooperates with the cyclin subunit to tune protein dynamics and promote catalytic activity, and 3) how dynamic conformational changes in Cdk2 control access of small-molecule ligands to allosteric pockets in the kinase. The insights from this work will fundamentally advance our understanding of allosteric control mechanisms in proteins, and help set the stage for the design of advanced allosteric therapeutics that effectively harness allostery to modulate kinase function.

项目摘要该项目达到了一个新的范式，以了解蛋白激酶的会议动态如何驱动它们的催化激活，并提供了一个重要的案例研究，以了解如何命令蛋白质动力用变构小分子人为地控制激酶活性。蛋白激酶是一个大量的信号蛋白，可以控制所有人的细胞生长和增殖真核细胞。激酶的行为就像分子开关一样，在催化活性的“ on”和不活跃的“关闭”状态以严格控制的方式带来了细胞生理学的规定变化。这严格的调节控制在癌症中被广泛破坏，并针对异常的激酶活性以较小分子药物现在是许多癌症治疗的重要组成部分。晶体结构为我们提供了激酶开关状态和关闭状态的静态图片，但它们很少告诉我们关于这些状态之间的蛋白质如何过渡，这是一个固有的动态过程。的本质这些动态过渡及其在疾病中的扰动方式仍然很大程度上晦涩难懂，这一事实已有阻碍了我们设计变构疗法的能力，通过模仿他们自然控制机制。相反，现有的激酶抑制剂通过与高度组成的活性结合来起作用站点，一种行动方式，使其选择性不佳。该项目的目的是使用提供完善的实验方法的组合有关蛋白质动力学的信息，以确定不同构象之间的激酶如何过渡状态，并了解如何通过小分子药物将激酶动态命令为人为调节激酶功能。使用核磁共振和光谱法应用于细胞周期蛋白依赖性激酶CDK2是细胞周期进程的关键调节剂，我们的目的是揭示1）变构耦合将细胞周期蛋白结合到CDK2激活的机制，2）磷酸化如何与细胞周期蛋白合作亚基调整蛋白质动力学并促进催化活性，3）动态会议如何变化 CDK2控制小分子配体进入激酶中的变质袋。这项工作的见解从根本上讲，我们将提高我们对蛋白质变构控制机制的理解，并帮助设定高级变构疗法设计的阶段，该疗法有效利用变构调节激酶功能。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Allostery governs Cdk2 activation and differential recognition of CDK inhibitors.

DOI：
10.1038/s41589-020-00725-y
发表时间：
2021-04
期刊：
Nature chemical biology
影响因子：
14.8
作者：
Majumdar A;Burban DJ;Muretta JM;Thompson AR;Engel TA;Rasmussen DM;Subrahmanian MV;Veglia G;Thomas DD;Levinson NM
通讯作者：
Levinson NM