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.

项目摘要/摘要 我们目前对体外蛋白质折叠的理解是由于数十年的实验和计算引起的 提供了蛋白质结构的高分辨率表征的研究，折叠原理的识别， 和折叠算法的开发。但是，蛋白质通常参与新的和意外的功能 体内病理行为。因为蛋白质过程涉及一个大的相互作用网络 强烈依赖于环境，了解细胞内蛋白质的工作方式需要了解蛋白质 体内结构，稳定性和动力学。虽然有证据表明细胞环境伴有蛋白质行为 半个多世纪以前出现了，我们仍然有关于这些影响的基本信息 关于蛋白质特性的合作细胞相互作用。知识的差距在很大程度上归因于弱者 相互作用在细胞环境中的瞬时性质以及与研究蛋白质功能相关的挑战 活细胞。由于细胞和生物中的蛋白质是连续相互作用的，因此这种限制是关于这种局限性的 使用其他生物分子，这可能会破坏蛋白质正确折叠和组装的能力，并导致 功能丧失和最终疾病。为了解决这些差距，我们的研究小组利用开创性 体内光谱微观显微镜法，结合功能性生化测定，体外生物物理 光谱和数值分析解决方案，以表征活细胞中蛋白质结构动力学和 组织。该平台将改变我们检查未开发的蛋白质复杂性和调节层的能力 在细胞和组织中，特别是：（1）经典的体外蛋白质原理会转化为细胞吗？有多准确 折叠理论和分子动力学模拟的细胞内预测？ （2）我们可以开发方法 可视化新陈代谢和相关代谢蛋白结构动力学的空间分布 细胞？ （3）生物体的热适应和适应性如何改变稳定性，折叠和 分化组织中蛋白质的聚集？总体而言，这项工作将使人们对 开发过程中细胞内部的重塑，环境应激和疾病有助于蛋白质 稳态。解开这些相互作用将改善我们对必需品的分子级别的理解 人类健康和疾病的过程。