Functional Protein Conformations and Dynamics via Transparent Window 1D & 2D Infrared Spectroscopy

通过透明窗口 1D 观察功能性蛋白质构象和动力学

• 批准号: 10552386
• 负责人: Megan Corrine Thielges
• 金额: $ 54.6万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-05 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552386
• 关键词: Adopted; Area; Binding; Biochemical; Biochemistry; Biological Process; Biology; Biophysics; Cells; Chemistry; Communication; Coupling; Development; Dimensions; Entropy; Frequencies; Health; Heterogeneity; Human; Label; Medicine; Methodology; Methods; Modeling; Modernization; Molecular Conformation; Molecular Machines; Motion; Population Dynamics; Postdoctoral Fellow; Process; Protein Conformation; Protein Dynamics; Proteins; Reaction; Research; Research Personnel; Role; Science; Side; Signal Transduction; Site; Solvents; Specificity; Spectrum Analysis; Structure; System; Training; Tumor Suppression; drug metabolism; infrared spectroscopy; multidisciplinary; programs; protein function; temporal measurement; undergraduate student; virtual

Project Summary/Abstract Proteins are the molecular machines of a cell that orchestrate virtually all processes. Like a macroscopic machine, their function requires that they move to adopt different states. Thus, a modern view of protein biophysics has expanded from the structure-function paradigm to include dynamics - population of ensembles of protein states (conformational heterogeneity) and their interconversion. Fully delineating the dynamics that underlie function is however complicated by the immense complexity of proteins, due to both their large size with spatial heterogeneity of their chemistry and the broad timescales over which states may interconvert, ranging from large-scale processes such as aggregation that occur over days to years to the picosecond fluctuations of side chains and solvent. Among these scales, the local small-scale changes in proteins that involve rapidly interconverting states are perhaps among the most challenging to characterize and least well understood. However, such motions are argued to be central to many aspects of protein function, such as main contributors to the entropy of reactions, allosteric communication within domains, catalytic and binding specificity, et al. This research program is directed at developing rigorous methodologies to advance understanding of fundamental protein biophysics in important biological processes. We are developing infrared (IR) spectroscopy as an approach with inherently high spatial and temporal resolution for accessing all involved conformational ensembles and dynamics. By incorporating vibrational groups with frequencies within a transparent window of protein IR spectra we avoid the complexity of spectral congestion to enable inspection of single vibrational modes at local sites anywhere throughout proteins. We are combining modern approaches in biochemistry for selective labeling with state-of-the-art methods in multidimensional spectroscopy to provide rigorous analysis of frequency heterogeneity, coupling, and dynamics. The application and development of new biochemical and spectroscopic methods should elucidate the biophysical foundations of protein function with unprecedented detail, providing information with promise to buttress advances in broad areas of biology and medicine. In addition, through execution of this project, undergraduate, graduate, and postdoctoral researchers will be broadly trained in multidisciplinary science, strengthening the nation’s scientific workforce.

项目概要/摘要 蛋白质是细胞的分子机器，几乎协调所有过程。就像宏观的 机器，它们的功能要求它们采取不同的状态。因此，蛋白质的现代观点 生物物理学已经从结构-功能范式扩展到包括动力学——系综种群 蛋白质状态（构象异质性）及其相互转换。充分描绘了动态 然而，蛋白质的巨大复杂性使其基础功能变得复杂，因为它们的尺寸很大 由于其化学的空间异质性以及状态可能相互转换的广泛时间尺度， 范围从大规模过程（例如几天到几年内发生的聚合）到皮秒 侧链和溶剂的波动。在这些尺度中，蛋白质的局部小尺度变化 涉及快速相互转换的状态可能是最具挑战性且最不好描述的状态 明白了。然而，此类运动被认为是蛋白质功能许多方面的核心，例如 反应熵、域内变构通讯、催化和结合的主要贡献者 特异性等。 该研究计划旨在开发严格的方法来增进对 重要生物过程中的基础蛋白质生物物理学。我们正在开发红外 (IR) 光谱 作为一种具有固有的高空间和时间分辨率的方法，用于访问所有涉及的构象 系综和动力学。通过将振动群与频率合并在透明窗口内 蛋白质红外光谱我们避免了光谱拥塞的复杂性，从而能够检查单一振动模式 在整个蛋白质的任何地方的局部位点。我们正在结合生物化学的现代方法来选择性地 使用多维光谱中最先进的方法进行标记，以提供严格的频率分析 异质性、耦合性和动态性。新型生化、光谱技术的应用与发展 方法应该以前所未有的细节阐明蛋白质功能的生物物理基础，提供 有望支持生物学和医学广泛领域进步的信息。此外，通过 在该项目的执行过程中，本科生、研究生和博士后研究人员将接受广泛的培训 多学科科学，加强国家科学队伍。