NMR Approaches for Structural Studies of Large Proteins and Protein Complexes

用于大蛋白质和蛋白质复合物结构研究的 NMR 方法

• 批准号: 7265738
• 负责人: GERHARD WAGNER
• 金额: $ 35.6万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7265738
• 关键词: Address; Amino Acid Motifs; Amino Acid Sequence; Attention; Behavior; Biological Process; Carbon; Cell physiology; Class; Collaborations; Complex; Computational Technique; Computer software; Crowding; Crystallography; Data; Data Analyses; Data Set; Development; Dimensions; Disease; Drug Design; Entropy; Event; Evolution; Flavoring; Funding; Gene Expression Regulation; Goals; Grant; Hand; Individual; Label; Laboratories; Ligands; Ligase; Measurement; Measures; Methane; Methods; Molecular; Mono-S; NMR Spectroscopy; Numbers; Oxygenases; Peptide Sequence Determination; Performance; Physics; Physiologic pulse; Positioning Attribute; Preparation; Procedures; Process; Proteins; Pulse taking; Recycling; Relaxation; Research; Residual state; Resolution; Sampling; Side; Signal Pathway; Signal Transduction; Solubility; Solutions; Structure; System; Techniques; Technology; Testing; Time; Work; base; computerized data processing; data acquisition; design; enzyme mechanism; improved; insight; instrument; method development; molecular dynamics; new technology; novel; novel strategies; optimal control theory; polypeptide; programs; protein protein interaction; reconstruction; research study; restraint; small molecule; theories

This research component has the overall goal to develop and improve methods for characterizing large proteins and protein complexes. Protein interactions are crucial for many biological processes, such as cellular switches, signaling mechanisms, enzyme regulation, gene expression and development. Much is known about structures of tight complexes from crystallography and NMR spectroscopy. However, information on weak complexes is rather sparse due to the limitations of current technologies. This is in contrast to the fact that many protein interactions must be weak and transient, for example to rapidly turn signaling pathways on and off, or to recycle cellular proteins. Furthermore, protein complexes represent an underutilized class of targets for design of drugs against diseases. NMR has unique capabilities for defining structures and states of large proteins and protein complexes that cannot be obtained with crystallography. Major improvements of NMR hardware have recently become available that are only beginning to be efficiently utilized. New ideas of spin physics, control theory, sampling strategies, signal processing, or data analysis are being developed, together with new strategies for sample preparation. All this promises major advances of NMR spectroscopy with large proteins and protein complexes. Here we propose to develop new approaches to make optimal use of modern NMR hardware to gain new insights into structures of large proteins and protein complexes. This will require abandoning some of the traditional procedures for data acquisition and require new data processing methods. This grant has spearheaded such approaches in the past, and similar efforts have appeared in several other laboratories. We propose research towards two specific aims: Aim 1. Develop new NMR experiments for characterization of large proteins systems. The experiments proposed employ heavily non-uniform sampling methods, coherence co-evolution procedures, and they are geared towards optimum use of high-field instruments and use optimum control theory for pulse sequence design. Aim 2. Approaches for characterizing protein complexes. This includes new co-expression methods for facilitating NMR studies of complexes, development of new protein tags to improve the solution behavior of complexes, and NMR and computational techniques for defining the arrangement of proteins in tight and in weak complexes.

该研究部分的总体目标是开发和改进表征大型 蛋白质和蛋白质复合物。蛋白质相互作用对于许多生物过程至关重要，例如 细胞开关、信号传导机制、酶调节、基因表达和发育。很多是 从晶体学和核磁共振波谱中了解紧密配合物的结构。然而， 由于当前技术的限制，关于弱复合体的信息相当稀疏。这是在 与此相反，许多蛋白质相互作用必须是微弱且短暂的，例如快速转变 信号通路的开启和关闭，或回收细胞蛋白质。此外，蛋白质复合物代表 设计抗疾病药物时未充分利用的靶标类别。 NMR 具有定义大蛋白质和蛋白质复合物的结构和状态的独特功能 这是晶体学无法获得的。核磁共振硬件的重大改进最近已成为 可用的资源才刚刚开始被有效利用。自旋物理、控制论、采样的新思想 正在开发策略、信号处理或数据分析，以及样本的新策略 准备。所有这些都预示着大蛋白质和蛋白质的核磁共振波谱学将取得重大进展 复合物。 在这里，我们建议开发新方法来充分利用现代 NMR 硬件来获得新的 深入了解大型蛋白质和蛋白质复合物的结构。这将需要放弃一些 传统的数据采集程序需要新的数据处理方法。这笔补助金有 过去曾带头采用此类方法，其他几个实验室也出现了类似的努力。 我们建议针对两个具体目标进行研究： 目标 1. 开发新的 NMR 实验来表征大型蛋白质系统。实验 提出采用严重不均匀的采样方法、一致性共同进化程序，它们是 旨在优化使用高场仪器并使用脉冲序列的优化控制理论 设计。 目标 2. 表征蛋白质复合物的方法。这包括新的共表达方法 促进复合物的 NMR 研究，开发新的蛋白质标签以改善复合物的溶液行为 复合物，核磁共振和计算技术来定义蛋白质的紧密排列 弱复合物。