Single Molecule Studies to Dissect Allosteric Communication Networks in Protein Kinase A

单分子研究剖析蛋白激酶 A 中的变构通讯网络

• 批准号: 1715572
• 负责人: Rodrigo Maillard
• 金额: $ 29.96万
• 依托单位: Georgetown University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1715572&HistoricalAwards=false
• 关键词: Single; Molecule; Studies; Dissect; Allosteric

Signaling proteins are  the core building blocks in the transmission of biological signals, allowing cells to communicate between the extracellular environment and the cellular interior.  Understanding how signaling proteins operate has important and broad implications, from defining how biological systems detect and transduce signals, to reengineering protein function for biotechnological applications, to identifying new binding sites for allosteric inhibitors. The cAMP-dependent Protein Kinase A (PKA) is a major signaling protein in all eukaryotic cells that incorporates structural and functional elements seen in most signaling protein complexes, including a modular domain organization, ligand binding cooperativity, signaling through flexible linker domains, and ligand-induced conformational changes. This protein will be used as a model system to quantitatively define how biological signals are transduced from one domain to another to regulate and activate protein function. This project will provide research opportunities to undergraduate and graduate students at the interface of biology, physics and computational  modeling.This project will use single-molecule biophysical techniques in combination with molecular dynamic simulations to investigate the mechanism of communication between different structural elements of PKA. The goal of this project is to determine how such communication mechanisms are established in different biological states (apo, active and  inactive  states), and how cAMP binding activates the protein. It is expected that the tools and methods developed from these studies will have widespread applications to advance knowledge in a variety of related and unrelated kinases that display modular architectures, such as cGMP-dependent protein kinase involved in vasodilation and  platelet activation, or  protein kinase C involved in cell migration, differentiation, and apoptosis. Lastly, the interdisciplinary nature of this project will provide unique research opportunities to graduate and undergraduate students from different backgrounds.

信号蛋白是生物信号传递的核心组成部分，允许细胞在细胞外环境和细胞内部之间进行通信。 了解信号蛋白如何运作具有重要而广泛的意义，从定义生物系统如何检测和识别信号，到为生物技术应用重新设计蛋白质功能，再到识别变构抑制剂的新结合位点。 cAMP依赖性蛋白激酶A（PKA）是所有真核细胞中的主要信号蛋白，其结合了在大多数信号蛋白复合物中可见的结构和功能元件，包括模块结构域组织、配体结合协同性、通过柔性接头结构域的信号传导以及配体诱导的构象变化。 该蛋白质将被用作模型系统，以定量定义生物信号如何从一个结构域转导到另一个结构域，以调节和激活蛋白质功能。 本计划将提供本科生及研究生在生物学、物理学及计算模型的研究机会，利用单分子生物物理技术结合分子动力学模拟，研究PKA不同结构元件之间的通讯机制。 该项目的目标是确定这种通信机制如何在不同的生物状态（apo，活性和非活性状态）中建立，以及cAMP结合如何激活蛋白质。 预计从这些研究中开发的工具和方法将具有广泛的应用，以推进显示模块化结构的各种相关和不相关激酶的知识，例如参与血管舒张和血小板活化的cGMP依赖性蛋白激酶，或参与细胞迁移、分化和凋亡的蛋白激酶C。 最后，该项目的跨学科性质将为来自不同背景的研究生和本科生提供独特的研究机会。

项目成果

Activation of PKA via asymmetric allosteric coupling of structurally conserved cyclic nucleotide binding domains

• DOI: 10.1038/s41467-019-11930-2
• 发表时间: 2019-09-04
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Hao, Yuxin;England, Jeneffer P.;Maillard, Rodrigo A.
• 通讯作者: Maillard, Rodrigo A.

Switching of the folding-energy landscape governs the allosteric activation of protein kinase A

• DOI: 10.1073/pnas.1802510115
• 发表时间: 2018-08-07
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: England, Jeneffer P.;Hao, Yuxin;Maillard, Rodrigo A.
• 通讯作者: Maillard, Rodrigo A.

Mechanical unfolding of a knotted protein unveils the kinetic and thermodynamic consequences of threading a polypeptide chain

• DOI: 10.1038/s41598-020-66258-5
• 发表时间: 2020-06
• 期刊: Scientific Reports
• 影响因子: 4.6
• 作者: M. Rivera;Yuxin Hao;Rodrigo A. Maillard;M. Báez