Protein Structural Dynamics in Living Cells

项目摘要/摘要 我们目前对蛋白质体外折叠的理解是几十年的实验和计算的结果 研究提供了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 以及体内的病理行为。因为蛋白质过程涉及一个巨大的相互作用网络， 强烈依赖于环境，了解蛋白质在细胞内的工作方式需要蛋白质的知识 体内的结构、稳定性和动力学。而有证据表明细胞环境扰乱了蛋白质的行为 出现在半个多世纪前，我们对这些影响的基本信息仍然有限 蛋白质性质上的协同细胞相互作用。知识的差距在很大程度上可归因于弱者 细胞环境中相互作用的瞬变性质和与研究蛋白质功能相关的挑战 活细胞。这一限制令人担忧，因为细胞和生物体中的蛋白质不断地相互作用 与其他生物分子一起，这可能会扰乱蛋白质正确折叠和组装的能力，并导致 丧失功能，最终导致疾病。为了解决这些差距，我们的研究小组利用开创性的 体内光谱显微镜方法，结合功能生化分析，体外生物物理 光谱学和数值分析解决方案，以表征活细胞和 纸巾。这个平台将改变我们检查未被探索的蛋白质复杂性和调控层的能力 在细胞和组织中，具体地说：(1)经典的体外蛋白质原理是否适用于细胞？这些数据有多准确？ 折叠理论和分子动力学模拟的细胞内预测？(2)我们能否开发方法来 可视化生活中新陈代谢的空间分布和相关的代谢蛋白结构动力学 细胞？(3)生物的热适应和驯化如何改变稳定性、折叠和 蛋白质在分化组织中的聚集？总体而言，这项工作将有助于更好地理解 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。 动态平衡。解开这些相互作用将提高我们对本质的分子水平的理解 人类健康和疾病的过程。

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.