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.

项目概要/摘要 我们目前对体外蛋白质折叠的理解是基于数十年的实验和计算 研究提供了蛋白质结构的高分辨率表征、折叠原理的识别、 和折叠算法的开发。然而，蛋白质经常参与新的和意想不到的功能 和体内病理行为。因为蛋白质过程涉及一个庞大的相互作用网络 强烈依赖于环境，了解蛋白质在细胞内如何工作需要蛋白质知识 结构、稳定性和体内动力学。虽然有证据表明细胞环境会扰乱蛋白质行为 半个多世纪前出现，我们对这些影响的基本信息仍然有限 细胞间的协同相互作用对蛋白质特性的影响。知识差距很大程度上归因于薄弱 细胞环境中相互作用的瞬时性质以及与研究蛋白质功能相关的挑战 活细胞。这种限制令人担忧，因为细胞和生物体中的蛋白质不断相互作用 与其他生物分子相互作用，这可能会破坏蛋白质正确折叠和组装的能力，并导致 功能丧失并最终导致疾病。为了解决这些差距，我们的研究小组利用了突破性的技术 体内光谱显微镜方法，结合功能生化测定、体外生物物理 光谱学和数值分析解决方案，用于表征活细胞中的蛋白质结构动力学 组织。该平台将改变我们检查未探索的蛋白质复杂性和调控层的能力 在细胞和组织中，特别是：（1）经典的体外蛋白质原理是否可以转化为细胞？准确度如何 折叠理论和分子动力学模拟的细胞内预测？ (2) 我们能否开发方法 可视化代谢的空间分布和相关的代谢蛋白质结构动态 细胞？ (3) 生物体的热适应和驯化如何改变稳定性、折叠和 分化组织中蛋白质的聚集？总的来说，这项工作将让人们更好地理解如何 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形成 体内平衡。解开这些相互作用将提高我们对基本物质的分子水平理解 人类健康和疾病的过程。