Protein Structural Dynamics in Living Cells

活细胞中的蛋白质结构动力学

• 批准号: 10712991
• 负责人: Caitlin Davis
• 金额: $ 41.88万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712991
• 关键词: Acclimatization; Address; Algorithms; Behavior; Biochemical; Biological Assay; Biophysics; Cell physiology; Cells; Development; Disease; Environment; Failure; Health; Human; In Vitro; Knowledge; Metabolic; Metabolism; Methods; Molecular; Nature; Organism; Pathologic; Pathologic Processes; Process; Property; Proteins; Regulation; Research; Resolution; Spatial Distribution; Spectrum Analysis; Stress; Technology; Tissue Differentiation; Tissues; Translating; Visualization; Work; imaging approach; improved; in vivo; loss of function; molecular dynamics; protein aggregation; protein folding; protein function; protein structure; proteostasis; theories

Project Summary/Abstract Our current understanding of in vitro protein folding is due to decades of experimental and computational research that provided high-resolution characterization of protein structure, identification of folding principles, and development of folding algorithms. However, proteins often participate in new and unexpected functional and pathological behaviors in vivo. Because protein processes involve a large network of interactions that strongly depend on the environment, understanding how proteins work inside cells requires knowledge of protein structure, stability, and dynamics in vivo. While evidence that the cellular environment perturbs protein behaviors emerged over half a century ago, we still have limited fundamental information about the effects of these cooperative cellular interactions on protein properties. The gap in knowledge is largely attributable to the weak transient nature of interactions in the cellular milieu and challenges associated with studying protein functions in living cells. This limitation is concerning because proteins in cells and organisms are continuously interacting with other biomolecules, which may disrupt the ability of a protein to fold and assemble properly and results in loss of function and eventually disease. To address these gaps, our research group leverages groundbreaking in vivo spectro-microscopy methods, in combination with functional biochemical assays, in vitro biophysical spectroscopy, and numerical analysis solutions to characterize protein structural dynamics in living cells and tissues. This platform will transform our ability to examine unexplored layers of protein complexity and regulation in cells and tissues, specifically: (1) Do classic in vitro protein principles translate to cells? How accurate are the in-cell predictions of folding theory and molecular dynamics simulations? (2) Can we develop methods to visualize the spatial distribution of metabolism and associated metabolic protein structural dynamics in living cells? (3) How does thermal adaptation and acclimation by organisms change the stability, folding, and aggregation of proteins in differentiated tissues? Overall, this work will lead to a greater understanding of how remodeling of the cell interior during development, environmental stress, and disease contributes to protein homeostasis. Unraveling these interactions will improve our molecular-level understanding of essential processes in human health and disease.

项目总结/摘要 我们目前对体外蛋白质折叠的理解是由于几十年的实验和计算 提供蛋白质结构的高分辨率表征，折叠原理的鉴定， 和折叠算法的发展。然而，蛋白质经常参与新的和意想不到的功能， 和病理行为。因为蛋白质过程涉及一个巨大的相互作用网络， 由于蛋白质强烈依赖于环境，因此了解蛋白质在细胞内的工作方式需要了解蛋白质 结构、稳定性和体内动力学。虽然细胞环境干扰蛋白质行为的证据 虽然这些技术已经出现了半个多世纪，但我们对这些技术的影响的基本信息仍然有限。 协同细胞相互作用对蛋白质性质的影响。知识上的差距主要是由弱者造成的 细胞环境中相互作用的瞬时性质以及与研究细胞中蛋白质功能相关的挑战 活细胞这种限制是令人担忧的，因为细胞和生物体中的蛋白质不断相互作用 这可能会破坏蛋白质正确折叠和组装的能力， 功能丧失最终导致疾病为了解决这些差距，我们的研究小组利用了开创性的 体内光谱显微镜方法，结合功能生化测定，体外生物物理 光谱和数值分析解决方案，以表征活细胞中的蛋白质结构动力学， 组织中这个平台将改变我们研究蛋白质复杂性和调控的未探索层的能力 在细胞和组织中，具体而言：（1）经典的体外蛋白质原理是否转化为细胞？如何准确的是， 折叠理论和分子动力学模拟的细胞内预测？(2)我们能开发出 可视化的空间分布的代谢和相关的代谢蛋白质结构动力学在生活 细胞？(3)生物体的热适应和驯化如何改变蛋白质的稳定性、折叠和 分化组织中蛋白质的聚集？总的来说，这项工作将导致更好地了解如何 在发育、环境压力和疾病过程中细胞内部的重塑有助于蛋白质的合成。 体内平衡解开这些相互作用将提高我们的分子水平的理解， 人类健康和疾病的过程。