Intrinsic Disorder, Energetic Coupling and Allostery

本征无序、能量偶联和变构目前的项目研究蛋白质本征无序(ID)在蛋白质转录因子家族的信号传递中的作用，这一过程被称为变构。通过测量一些天然存在的糖皮质激素受体(GR)转录因子的稳定性和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的蛋白质中的变构信号提供了一个框架。由此产生的实验数据将用于构建变构的定量预测模型。拟议活动的智力价值有两个方面。首先，这些研究首次使用生物物理研究和活细胞功能分析对ID介导的变构进行了系统分析。其次，这些研究挑战了最近发展的系综变构模型，该模型旨在根据结构中合作元素的内在稳定性和它们之间的相互作用能量来定量表征变构。因此，这项研究代表了对ID功能的定性和主要是推测的模型的进步，并提供了关于蛋白质如何以及为什么使用内在紊乱的第一批定量描述之一。该项目的更广泛的影响是双重的，并侧重于研究、教育和这两个活动之间的桥梁。首先，这项研究的主要目标是通过实验确定GR中存在的变构控制。然而，GR与雌激素(ER)、孕激素(PR)、雄激素(AR)和维生素D(VDR)受体具有相同的结构，所有这些受体都在激素依赖的细胞信号和调节中发挥重要作用。因此，从当前研究中获得的见解将直接影响对这些其他系统的理解。其次，这项研究的一个关键目标是推导出一个经过模拟和验证的量化模型。作为之前两项NSF提案的一部分，首席研究员开发了大量以计算机为基础的课程工作，重点是动态生物系统的建模。作为当前研究的一部分开发的模型将直接整合到约翰·霍普金斯大学的研究生和本科生课程中，因此不仅将在生物学学生的教育中发挥至关重要的作用，还将显著扩大生物学学生的规模？获得计算方法和技术。

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.