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

• 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
• 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

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 该项目为蛋白激酶的构象动力学如何驱动提供了一个新的范式 它们的催化激活，并提供了如何控制蛋白质动力学的重要案例研究 用变构小分子人为地控制激酶活性。 蛋白激酶是一个信号蛋白大家族，可控制所有细胞的生长和增殖。 真核细胞。激酶的行为类似于分子开关，在催化活性“开启”和“开启”之间转换 以严格控制的方式非活动的“关闭”状态，以引起细胞生理学的规定变化。这 严格的监管控制在癌症中被广泛破坏，并以小分子靶向异常激酶活性 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启和关闭状态的静态图片，但它们告诉我们的信息很少 关于蛋白质如何在这些状态之间转变，这是一个固有的动态过程。的性质 这些动态转变以及它们在疾病中如何受到干扰仍然在很大程度上不清楚，这一事实已被证实 阻碍了我们设计变构疗法的能力，通过模仿激酶将激酶切换到关闭状态 自然控制机制。相反，现有的激酶抑制剂通过与高度保守的活性物质结合来发挥作用。 网站，这种行动方式使他们缺乏选择性。 该项目的目标是结合使用提供互补的实验方法 有关蛋白质动力学的信息，以确定激酶如何在不同构象之间转变 状态，并了解小分子药物如何人为地控制激酶动力学 调节激酶功能。使用核磁共振和光学光谱法应用于细胞周期蛋白 依赖激酶 Cdk2 是细胞周期进程的关键调节因子，我们的目标是揭示 1) 变构耦合 将细胞周期蛋白结合与 Cdk2 激活联系起来的机制，2) 磷酸化如何与细胞周期蛋白配合 亚基调节蛋白质动力学并促进催化活性，以及​​3）动态构象变化如何 Cdk2 控制小分子配体进入激酶中的变构袋。这项工作的见解 将从根本上增进我们对蛋白质变构控制机制的理解，并帮助设定 设计先进变构疗法的阶段，可有效利用变构来调节激酶 功能。

项目成果

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