Low-complexity domain protein molecular structure, conformational dynamics, and inter-protein interactions in human health and disease

人类健康和疾病中的低复杂性域蛋白质分子结构、构象动力学和蛋白质间相互作用

• 批准号: 10275947
• 负责人: Dylan Thomas Murray
• 金额: $ 37.21万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10275947
• 关键词: Amino Acid Sequence; Amino Acids; Antibodies; Behavior; Biochemical; Biological; Biological Assay; Biological Process; Cells; Characteristics; Code; Coupled; Cryoelectron Microscopy; Cytoplasmic Granules; Cytoskeleton; DNA Sequence Alteration; Dementia; Development; Disease; Health; Human; Human Genome; In Vitro; Intermediate Filaments; Investigation; Knowledge; Life; Link; Malignant Neoplasms; Methodology; Microscopic; Modeling; Molecular; Molecular Conformation; Molecular Structure; Motor Neuron Disease; Muscular Dystrophies; Neurodegenerative Disorders; Nuclear Magnetic Resonance; Organelles; Pathogenicity; Play; Point Mutation; Post-Translational Protein Processing; Process; Property; Protein Chemistry; Protein Dynamics; Protein Engineering; Proteins; RNA; RNA metabolism; Research; Role; Spectrum Analysis; System; Tertiary Protein Structure; Time; Work; base; biological systems; biophysical analysis; biophysical techniques; clinical biomarkers; improved; in vivo; protein expression; protein protein interaction; self assembly; small molecule therapeutics; tool

Project Summary/Abstract The dynamic assembly of biomolecules within a living cell is vital for the spatial and temporal organization of biological function. In forming RNA granule membraneless organelles and intermediate filaments in the cell cytoskeleton, cells leverage the self-assembly properties of protein sequences with reduced amino acid diversity. These low complexity protein domains have only recently come to light as essential players in these processes. Thirty percent of the proteins coded by the human genome contain a domain of this type, highlighting the central importance of these sequences for life. In humans, pathogenic genetic mutations and altered expression levels, in addition to functional post-translational modifications and protein-protein interactions, modulate the assembly processes of these proteins. Linked by the common involvement of low complexity domain proteins, this proposal outlines two lines of research focused on a fundamental mechanistic understanding of how the proteins that compose RNA granules and intermediate filament networks assemble to achieve the macroscopic behavior observed in living cells. A multifaceted biophysical approach employing cutting-edge nuclear magnetic resonance and cryo-electron microscopy will allow characterization of the molecular structure and conformational dynamics of these proteins in biologically relevant assemblies. These biophysical studies will be coupled with other spectroscopies, biochemical assays, and protein engineering to form more comprehensive models of how low complexity domain proteins assemble temporally and spatially. The results of these efforts will provide a mechanistic description of how these assembly processes and their associated control mechanisms are modulated by point mutations and altered protein expression levels linked to motor neuron disease, dementia, muscular dystrophy, and cancer. The in vitro work proposed here will provide detailed and testable models regarding the function of in vivo biological assemblies involved in RNA metabolism and the cell cytoskeleton. In the broader context of human health, the molecular characterizations of disease-relevant low complexity domain proteins and their interacting molecular partners will provide a base of knowledge useful for the exploration of these systems as clinical biomarkers and will also facilitate the development of antibody and small molecule therapeutics. Beyond the specific biological systems discussed in this proposal, the tools and methodologies employed here are expected to have applicability and impact on investigations of the thirty percent of the proteins in the human genome that contain a low complexity domain.

项目概要/摘要 活细胞内生物分子的动态组装对于空间和时间组织至关重要 的生物学功能。在细胞内形成RNA颗粒、无膜细胞器和中间丝 细胞骨架，细胞利用氨基酸减少的蛋白质序列的自组装特性 多样性。这些低复杂性蛋白质结构域直到最近才作为这些领域的重要参与者而被发现 流程。人类基因组编码的蛋白质中有百分之三十包含这种类型的结构域， 强调这些序列对生命的核心重要性。在人类中，致病性基因突变和 除了功能性翻译后修饰和蛋白质-蛋白质外，还改变了表达水平 相互作用，调节这些蛋白质的组装过程。通过低水平的共同参与联系起来 复杂性域蛋白质，该提案概述了侧重于基本机制的两条研究路线 了解组成 RNA 颗粒和中间丝网络的蛋白质如何组装 以实现在活细胞中观察到的宏观行为。采用多方面的生物物理方法 尖端核磁共振和冷冻电子显微镜将能够表征 这些蛋白质在生物学相关组件中的分子结构和构象动力学。这些 生物物理学研究将与其他光谱学、生化测定和蛋白质工程相结合，以 形成关于低复杂性域蛋白质如何在时间和空间上组装的更全面的模型。 这些努力的结果将为这些组装过程及其如何进行提供机械描述。 相关的控制机制由点突变和改变的蛋白质表达水平调节 运动神经元疾病、痴呆、肌营养不良和癌症。这里提出的体外工作将 提供有关 RNA 体内生物组装体功能的详细且可测试的模型 新陈代谢和细胞骨架。在更广泛的人类健康背景下，分子特征 疾病相关的低复杂性结构域蛋白及其相互作用的分子伙伴将提供基础 对于探索这些系统作为临床生物标志物有用的知识，也将促进 抗体和小分子疗法的开发。除了讨论的特定生物系统之外 该提案、此处使用的工具和方法预计将具有适用性和影响 对人类基因组中 30% 包含低复杂性结构域的蛋白质的研究。