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: 8072536
• 负责人: GERHARD WAGNER
• 金额: $ 26.58万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8072536
• 关键词: Address; Amino Acid Motifs; Amino Acid Sequence; Attention; Behavior; Biological Process; Carbon; Cell physiology; 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; Oxygenases; Peptide Sequence Determination; Performance; Physics; Physiologic pulse; Positioning Attribute; Preparation; Procedures; Process; Proteins; 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; meetings; method development; molecular dynamics; new technology; novel; novel strategies; optimal control theory; polypeptide; programs; protein complex; 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规模 蛋白质和蛋白质复合物。蛋白质相互作用对许多生物过程至关重要，例如 细胞开关、信号机制、酶调节、基因表达和发育。多问题 从晶体学和核磁共振光谱学中了解紧密复合物的结构。然而，在这方面， 由于现有技术的限制，关于弱络合物的信息相当稀少。这是 与许多蛋白质相互作用必须是弱的和瞬时的事实相反，例如为了快速地转变为 信号通路的开启和关闭，或者细胞蛋白质的再循环。此外，蛋白质复合物代表了 未充分利用的一类靶点，用于设计抗疾病药物。 核磁共振具有确定大蛋白质和蛋白质复合物的结构和状态的独特能力 这是晶体学无法获得的。最近，NMR硬件的主要改进已经成为 可用的资源才刚刚开始得到有效利用。自旋物理、控制论、采样的新思想 策略，信号处理或数据分析正在开发中，同时还有新的采样策略。 准备.所有这一切都预示着核磁共振光谱与大蛋白质和蛋白质的重大进展 配合物 在这里，我们建议开发新的方法，以最佳利用现代核磁共振硬件，以获得新的 深入了解大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。这笔赠款 在过去率先采取了这种方法，在其他几个实验室也出现了类似的努力。 我们建议研究两个具体目标： 目标1。开发新的NMR实验，用于表征大型蛋白质系统。实验 建议采用严重的非均匀采样方法，相干协同进化程序，他们是 以高场仪器的最佳使用为目标，并对脉冲序列使用最佳控制理论 设计 目标二。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，用于定义蛋白质的排列， 弱络合物