NMR Determination of Biomolecular Structural Dynamics Using Residual Dipolar Couplings Measured in Multiple Alignment Media

使用在多种对准介质中测量的残余偶极耦合进行生物分子结构动力学的 NMR 测定

• 批准号: 0615786
• 负责人: Joel Tolman
• 金额: $ 94.62万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0615786&HistoricalAwards=false
• 关键词: NMR; Determination; Biomolecular; Structural; Dynamics

The goal of this project is the development of residual dipolar coupling (RDC)-based NMR methods for the determination of biomolecular structure and dynamics. This approach will be based on the measurement of multiple sets of RDCs under conditions of different anisotropic solvent environments. Novel methods for modulating the nature of the anisotropic solvent environment will be developed in order to improve the efficiency of data acquisition. In particular, these studies will focus on the exploitation of the interference observed between aligning forces for composite, mechanically ordered media such as bacteriophage Pf1 particles embedded within a strained polyacrylamide gel matrix. The DIDC approach to the interpretation of RDCs, demonstrated by the PI for backbone amide N-H bonds, will be extended to the determination of side-chain conformation and dynamics. Furthermore, these multi-alignment RDC-based approaches will be adapted for application to the protein Neisseria meningitidis heme oxygenase, for which dynamics have been implicated as necessary for its function. These studies will serve as a test case for the ability to characterize in detail the structural dynamics of a moderately sized protein (24 kDa) using RDC-based methods. It is anticipated that these studies will lay a foundation for future NMR investigations and provide novel insights into the relationship between protein structural dynamics and function. This research will provide powerful tools for the characterization of mobility and structure of biomolecular systems and thus will have a far reaching impact on the ability to characterize the mechanical aspects of biomolecular function at atomic resolution. This project will also broadly impact undergraduate teaching and research in biomolecular NMR spectroscopy, a technique often inaccessible to undergraduates due to the expense and geographical concentration of high field NMR instrumentation. In order to increase undergraduate research opportunities in biomolecular NMR, one undergraduate summer research position has been established, primarily intended for students outside of the Johns Hopkins community. Sustained efforts are being made to stimulate collaboration and facilitate training of local biophysical and biochemical researchers such that they can employ state of the art biomolecular NMR spectroscopic techniques to advantage

该项目的目标是开发基于残留偶极耦合（RDC）的NMR方法，用于确定生物分子的结构和动力学。 该方法将基于在不同各向异性溶剂环境的条件下测量多组RDC。 将开发用于调制各向异性溶剂环境的性质的新方法，以提高数据采集的效率。 特别是，这些研究将集中在利用复合材料，机械有序的介质，如噬菌体Pf 1颗粒嵌入在一个应变的聚丙烯酰胺凝胶基质的对齐力之间观察到的干扰。 DIDC的方法来解释RDCs，证明了由PI的骨干酰胺N-H键，将扩展到侧链构象和动力学的测定。 此外，这些多对齐的基于RDC的方法将适用于应用于蛋白质脑膜炎奈瑟氏球菌血红素加氧酶，动力学已牵连其功能所必需的。 这些研究将作为一个测试案例，详细描述了一个中等大小的蛋白质（24 kDa）的结构动力学，使用基于RDC的方法的能力。 预计这些研究将为未来的NMR研究奠定基础，并为蛋白质结构动力学和功能之间的关系提供新的见解。 这项研究将为表征生物分子系统的流动性和结构提供强有力的工具，从而对以原子分辨率表征生物分子功能的机械方面的能力产生深远的影响。 该项目还将广泛影响生物分子NMR光谱学的本科教学和研究，由于高场NMR仪器的费用和地理集中，本科生通常无法获得这项技术。 为了增加本科生在生物分子核磁共振研究的机会，一个本科生暑期研究职位已经成立，主要是为学生以外的约翰霍普金斯社区。 我们正不断努力促进合作，并促进本地生物物理和生物化学研究人员的培训，使他们能够利用最先进的生物分子核磁共振光谱技术，