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.

该研究组成部分的总体目标是开发和改进表征大型的方法 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 关于晶体学和NMR光谱的紧密复合物结构已知。然而， 由于当前技术的局限性，有关弱复合物的信息相当稀少。这是 与许多蛋白质相互作用必须弱且短暂的事实形成鲜明对比，例如迅速转动 信号通路开关，或回收细胞蛋白。此外，蛋白质复合物代表 用于设计针对疾病的药物的靶标类别。 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 晶体学无法获得。 NMR硬件的重大改进已成为 可用的才开始有效地使用。旋转物理学，控制理论，抽样的新思想 正在制定策略，信号处理或数据分析以及样本的新策略 准备。所有这些有望用大蛋白和蛋白质的NMR光谱法的重大进展 复合物。 在这里，我们建议开发新的方法，以最佳使用现代NMR硬件以获得新的方法 对大蛋白质和蛋白质复合物结构的见解。这将需要放弃一些 传统的数据获取程序，需要新的数据处理方法。这笔赠款有 过去，这种方法率先进行，并且在其他几个实验室中也出现了类似的努力。 我们提出针对两个具体目标的研究： 目标1。开发新的NMR实验，以表征大蛋白质系统。实验 拟议采用严重的不均匀采样方法，相干共同进化程序，它们是 旨在最佳使用高场仪器，并将最佳控制理论用于脉冲序列 设计。 AIM 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，开发新蛋白质标签以改善溶液行为 复合物以及NMR和计算技术，用于定义紧密和中的蛋白质布置 弱复合物。