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Quantitative Description of Phosphorylation Effects on Disordered Protein Structure

磷酸化对无序蛋白质结构影响的定量描述

基本信息

批准号：
8940910
负责人：
STEVEN T WHITTEN
金额：
$ 33.56万
依托单位：
TEXAS STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8940910
关键词：
Affect Aging Alanine Amino Acid Substitution Amino Acids Biological Biological Models Biology Biophysics Charge Coupling Data Descriptor Development Dimensions Disease Electrostatics Exhibits Experimental Models Glycine Goals Homologous Gene Human Life Link Malignant Neoplasms Measurement Measures Methods Microscopic Modeling Molecular Molecular Conformation Molecular Models Monitor N-terminal Phosphorylation Phosphotransferases Positioning Attribute Process Proline Property Protein Dynamics Protein Region Protein p53 Proteins Radial Recombinants Role Signal Transduction Site Solutions Step Tests Structure Structure-Activity Relationship System Technology Testing Translating Tumor Suppressor Proteins Variant Work base light scattering molecular modeling polyproline protein structure protein structure function public health relevance research study role model

项目摘要

DESCRIPTION (provided by applicant): We have developed a method for characterizing intrinsically disordered protein (IDP) structures that we will use to establish a predictive and quantitative model that describes protein phosphorylation in disordered regions. Protein phosphorylation is used biologically as a mechanism to control many critical life processes and phosphorylation usually occurs within protein regions that are intrinsically disordered. Accordingly, quantitative descriptions and molecular models of phosphorylation effects on disordered protein structures are needed to understand the molecular basis of key aspects of development, aging, and disease. The method that we have developed to study IDPs is useful because it can analyze structural relationships in detail and with quantitative precision, linking microscopic residue-specific descriptors, such as intrinsic conformational propensities, to macroscopic global metrics like the hydrodynamic radius (Rh). For example, we demonstrated that the effects of glycine substitutions on Rh could be used to estimate per-residue polyproline II (PPII) propensities in disordered proteins. Our results also provided evidence that PPII propensities and charge effects on IDP structures are linked, indicating a possible correlation between PPII structure, which is a dominant conformation in disordered proteins, and phosphorylation effects, which are key regulators of IDP activity. For the studies that are proposed in this R-15 application, we will use the intrinsically disordered N-terminal region of th p53 tumor suppressor protein consisting of residues 1-93 as our experimental model system. Aberrant p53 activity has been associated with numerous human cancers. The objectives of this application are to test the ability of our method to extract structural detail from the p53(1-93) system, measure intrinsic PPII propensities for the common amino acid types, establish if intrinsic PPII propensities depend on nearest neighbor sequence details, and apply our technology for investigating phosphorylation mechanisms quantitatively by modeling the effects of charge and PPII propensities on IDP structure for comparison to experiments that will measure charge, phosphorylation, and PPII effects on Rh. The data measured will be fundamental for establishing a new and predictive approach for characterizing IDP structures, investigating their biological roles, and modeling how their biological activities are regulated. The goals of this application are part of our long-term objective of developing quantitative models of IDP structure/function relationships. The data from this study will advance IDP molecular biophysics by: 1) developing a method for measuring PPII propensities in disordered proteins, 2) establishing if local sequence details influence intrinsic PPII propensities, 3) quantifying coupling between charge and PPII effects on IDP structure, and 4) measuring quantitatively the effects of phosphorylation on IDP structure.

描述（由申请人提供）：我们已经开发了一种用于表征内在无序蛋白（IDP）结构的方法，我们将使用该方法建立描述无序区域中蛋白质磷酸化的预测和定量模型。蛋白质磷酸化在生物学上被用作控制许多关键生命过程的机制，并且磷酸化通常发生在本质上无序的蛋白质区域内。因此，需要对磷酸化对无序蛋白质结构的影响进行定量描述和分子模型，以了解发育，衰老和疾病的关键方面的分子基础。我们已经开发的研究IDP的方法是有用的，因为它可以详细分析结构关系，并具有定量精度，将微观残留物特异性描述符（如内在构象倾向）与宏观全局指标（如流体动力学半径（Rh））联系起来。例如，我们证明了甘氨酸取代对Rh的影响可用于估计无序蛋白质中的每残基聚脯氨酸II（PPII）倾向。我们的研究结果还提供了证据表明，PPII倾向和IDP结构上的电荷效应是联系在一起的，表明PPII结构，这是一个占主导地位的构象在无序的蛋白质，和磷酸化的影响，这是IDP活动的关键调节之间可能存在相关性。对于本R-15申请中提出的研究，我们将使用由残基1-93组成的p53肿瘤抑制蛋白的固有无序N-末端区域作为我们的实验模型系统。异常的p53活性与许多人类癌症有关。本申请的目的是测试我们的方法从p53（1-93）系统提取结构细节、测量常见氨基酸类型的内在PPII倾向、确定内在PPII倾向是否依赖于最近邻序列细节、并应用我们的技术，通过模拟电荷和PPII倾向对IDP结构的影响，定量研究磷酸化机制，与将测量电荷、磷酸化和PPII对Rh的影响的实验进行比较。测量的数据将是建立一个新的和预测性的方法来表征IDP结构，研究它们的生物学作用，并模拟它们的生物活性是如何调节的基础。该应用程序的目标是我们开发IDP结构/功能关系定量模型的长期目标的一部分。这项研究的数据将通过以下方式推进IDP分子生物物理学：1）开发一种测量无序蛋白质中PPII倾向的方法，2）确定局部序列细节是否影响内在PPII倾向，3）量化电荷和PPII对IDP结构的影响之间的耦合，以及4）定量测量磷酸化对IDP结构的影响。