The molecular determinants of surface-templated self-association of intrinsically disordered proteins

本质无序蛋白质表面模板自缔合的分子决定因素

• 批准号: 10715794
• 负责人: Peter J Chung
• 金额: $ 41.39万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-15 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10715794
• 关键词: Affinity; Behavior; Biological; Biological Models; Biophysics; Communities; Disease; Goals; Intention; Laboratories; Membrane; Methods; Microtubules; Modification; Molecular; Molecular Biology; Neurobiology; Neurodegenerative Disorders; Neurons; Organelles; Physical condensation; Physiological; Physiology; Predisposition; Protein Conformation; Proteins; Protocols documentation; Public Health; Recording of previous events; Research; Structure; Surface; Synaptic Vesicles; Techniques; alpha synuclein; insight; multidisciplinary; novel; programs; protein function; tau Proteins; tool

Project Summary The long-term goal of our research program is to utilize new techniques and paradigms to understand the molecular determinants of physiological and disease-relevant phenomena associated with intrinsically disordered proteins (IDPs), or proteins that do not fold into stable structures in solution. Our focus is the self- association of IDPs that occur on high-affinity surfaces (such as microtubules or organelle membranes). Surfaces can promote self-association by increasing the local IDP concentration and templating IDP conformations more susceptible to self-association. However, both IDPs and their respective high-affinity surfaces are subject to numerous cellular modifications, dramatically expanding the experimental parameter space necessary to precisely characterize this phenomenon. Understanding how self-association is controlled in this space could be central to understanding their function in physiology and disease. Thus, our laboratory will adapt novel tools beyond traditional molecular biology to recreate conditions in two model systems where this phenomenon occurs: Tau condensation on microtubules and α-synuclein multimerization on synaptic vesicle membranes. Not only does the PI have extensive expertise with biophysically characterizing these IDPs, but the importance of Tau and α-synuclein to neurobiology and neurodegenerative disease provide a rich history of experimental insights that can be applied towards this phenomenon. Combined, this expertise and background can be incorporated into the phenomenon of surface-templated self-association of these IDPs. Furthermore, we will establish protocols/methods that can be easily exported to study other IDPs that undergo surface-templated self- association, as well. Overall, our intention by precisely understanding phenomena associated with select IDPs is to create generalizable mechanisms by which other IDPs behave, eventually providing a rigorous framework that has explanatory and predictive power for these proteins. By undertaking the proposed research, we hope to transition our purely biophysics laboratory to an entirely multidisciplinary program that connects protein behavior to cellular phenomenon.

项目摘要 我们的研究计划的长期目标是利用新技术和范式来了解 与本质上有关的物理和疾病现象的分子决定剂 蛋白质无序蛋白（IDP）或蛋白质不折叠成溶液中稳定结构的蛋白质。我们的重点是自我 在高亲和力表面（例如微管或细胞器膜）上发生的IDP的关联。表面 可以通过增加局部IDP浓度和模板IDP构象来促进自我关联 容易自我关联。但是，IDP及其各自的高亲和力表面都需要 大量的细胞修饰，动态扩展了实验参数空间 精确地表征了这种现象。了解在这个领域如何控制自我关联的可能是 了解它们在生理和疾病中的功能的核心。那，我们的实验室将适应新颖的工具 除传统的分子生物学外，还可以在这种现象的两个模型系统中重现条件 发生：在微管上的Tau缩合和合成囊泡膜上的α-突触核蛋白多层化。不是 只有PI才具有广泛的专业知识，可以在生物物理上表征这些IDP，但是 TAU和α-突触核蛋白神经生物学和神经退行性疾病提供了丰富的实验史 可以应用于这种现象的见解。结合在一起，这种专业知识和背景可以是 纳入这些IDP的表面自我关联的现象中。此外，我们会的 建立可以轻松导出的协议/方法研究其他经历表面自我的IDP 协会。总体而言，我们的意图准确地理解与精选IDP相关的现象 是创建其他IDP行为的可推广机制，有时提供严格的框架 这具有这些蛋白质的剥夺和预测能力。通过进行拟议的研究，我们希望 将纯粹的生物物理学实验室转变为完全多学科的计划，该计划连接蛋白质行为 到细胞现象。