Intrinsic Disorder, Energetic Coupling and Allostery

内在紊乱、能量耦合和变构

• 批准号: 1330211
• 负责人: Vincent Hilser
• 金额: $ 58.73万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2017-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1330211&HistoricalAwards=false
• 关键词: Intrinsic; Disorder; Energetic; Coupling; Allostery

Intrinsic Disorder, Energetic Coupling and AllosteryThe current project investigates the role of protein intrinsic disorder (ID) in mediating signaling in the transcription factor family of proteins, a process known as allostery. By measuring the stability and DNA binding affinity for a number of naturally occurring variants of the glucocorticoid receptor (GR) transcription factor and comparing these values to the transcriptional activity in cells, this project provides a framework for understanding allosteric signaling in proteins containing ID. The resulting experimental data will be used to construct a quantitative, predictive model of allostery. The intellectual merits of the proposed activities are two-fold. First these studies provide the first systematic analysis of ID-mediated allostery using both biophysical studies and live cell assays of function. Second, these studies challenge a recently developed ensemble allosteric model designed to quantitatively characterize allostery in terms of the intrinsic stabilities of cooperative elements of structure and the interaction energies between them. As such, this research represents an advance over previous qualitative and largely speculative models for ID function, and provides one of the first quantitative descriptions of how and why proteins use intrinsic disorder.The broader impacts of the project are two-fold, and focus on research, education and the bridge between these two activities. First, the primary goal of the research is to experimentally determine the allosteric control present in GR. However, GR shares architecture with the estrogen (ER), progesterone (PR), androgen (AR), and vitamin D (VDR) receptors, all of which play a vital role in hormone-dependent cell signaling and regulation. As such, insights gained from the current research will directly impact understanding in these other systems. Second, a key objective of the research is to derive a quantitative model that is subject to simulation and validation. As part of two previous NSF proposals, the Principal Investigator has developed a significant amount of computer-based course work focuses on modeling of dynamic biological systems. The models developed as part of the current research will be directly integrated into the graduate and undergraduate curriculum at Johns Hopkins University, and thus will not only play a vital role in the education of biology students, it will significantly expand the biology students? access to computational methods and technologies.

固有障碍，能量耦合和变构的当前项目研究了蛋白质内在疾病（ID）在蛋白质转录因子家族中介导信号传导中的作用，这一过程称为变构。通过测量糖皮质激素受体（GR）转录因子的许多天然发生变体的稳定性和DNA结合亲和力，并将这些值与细胞中的转录活性进行比较，该项目为理解含有ID蛋白质的变构信号传导提供了一个框架。所得的实验数据将用于构建变构的定量，预测模型。拟议活动的智力优点是两个方面。首先，这些研究提供了使用生物物理研究和功能活细胞测定法进行ID介导的变构的首次系统分析。其次，这些研究挑战了最近开发的集合变构模型，旨在定量地以结构合作元素的内在稳定性以及它们之间的相互作用能量的固有稳定性来定量表征变构。因此，这项研究代表了以前的定性和很大程度上投机的ID功能模型的进步，并提供了关于蛋白质如何以及为什么使用内在障碍的最初定量描述之一。该项目的更广泛影响是两倍，并侧重于研究，教育和这两种活动之间的桥梁。首先，该研究的主要目标是实验确定GR中存在的变构控制。但是，GR与雌激素（ER），孕酮（PR），雄激素（AR）和维生素D（VDR）受体共享结构，所有这些受体在激素依赖性细胞信号传导和调节中都起着至关重要的作用。因此，从当前的研究中获得的见解将直接影响这些其他系统的理解。其次，研究的关键目的是得出受模拟和验证的定量模型。作为先前的两个NSF提案的一部分，主要研究人员已经开发了大量基于计算机的课程工作，重点是建模动态生物系统。作为当前研究的一部分开发的模型将直接纳入约翰·霍普金斯大学的研究生和本科课程，因此不仅将在生物学学生的教育中发挥至关重要的作用，而且会大大扩展生物学学生吗？访问计算方法和技术。