750 MHZ NMR SPECTROMETER

750 MHz 核磁共振波谱仪

• 批准号: 2284238
• 负责人: PETER Edwin WRIGHT
• 金额: $ 40万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-09-30 至 1995-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2284238
• 关键词: biomedical equipment purchase; biomedical equipment resource; magnetic field; nuclear magnetic resonance spectroscopy; nucleic acid structure; protein structure; structural biology; superconductivity

In recent years, high resolution NMR spectroscopy has developed into a second method, alongside X-ray crystallography, for determination of the 3D structures of biological macromolecules. Technological advances, both in the development of powerful multi-dimensional NMR experiments and of high-field (up to 14 Tesla) spectrometers, have steadily increased the molecular weight limits for NMR structural studies. However, many important problems in structural biology remain inaccessible using current techniques and spectrometers. There is clearly a need for both new experiments and higher field spectrometers. This proposal requests funding to support the purchase of a 17.6 Tesla (750 MHz 1H frequency) NMR spectrometer for studies of the structure and dynamics of proteins and nucleic acids. The biomolecular NMR group in the Department of Molecular Biology is currently staffed-by 6 senior personnel, 27 postdoctoral fellows, and 3 graduate students. Research activities include both development of new and improved NMR methods and applications of NMR to challenging problems of fundamental importance in structural biology. Many of our ongoing projects are pushing the limits of what is currently possible using existing NMR methodology and spectrometers. Higher magnetic fields give increased resolution and sensitivity, both of which are important in extending the NMR method to systems of higher molecular weight and greater complexity. Based on our experience with 500 and 600 MHz spectrometers, the increased sensitivity and dispersion of the 750 MHz instrument will have a significant impact on our ability to solve many problems. The increased dispersion will probably be most important for systems which cannot be labeled with 15N or 13C and for which homonuclear 1H NMR experiments must be used; examples of such systems include peptide models of protein folding intermediates, protein molten globule states, and DNA Holliday junctions. For some systems, such as multiple zinc finger proteins, protein-DNA complexes and RNA, both isotope labeling and the high dispersion of the 750 MHz spectrometer are likely to be of great importance. The increased sensitivity of the instrument will also be particularly beneficial for projects which are limited to concentrations of lmM or less due to problems of solubility or aggregation.

近年来，高分辨率NMR光谱学已经发展成为一种新的方法。 第二种方法，与X射线晶体学一起，用于确定 生物大分子的3D结构。技术进步，两者 在发展强大的多维核磁共振实验和 高场（高达14特斯拉）光谱仪，稳步增加了 NMR结构研究的分子量限制。但不少 结构生物学中的重要问题仍然无法利用当前的 技术和光谱仪。显然，需要两个新的 实验和更高场光谱仪。该提案要求提供资金 支持购买17.6特斯拉（750 MHz 1H频率）NMR 用于研究蛋白质结构和动力学的光谱仪， 核酸 生物分子核磁共振组在分子系 生物学目前的工作人员-由6名高级人员，27名博士后 研究员和3名研究生。研究活动包括 新的和改进的NMR方法的发展和NMR的应用， 结构生物学中具有根本重要性的挑战性问题。许多 我们正在进行的项目正在推动目前的极限， 可能使用现有的NMR方法和光谱仪。更高的磁 字段提供了更高的分辨率和灵敏度，这两者都是 重要的是将NMR方法扩展到更高分子的系统 重量和更大的复杂性。根据我们对500和600的经验 750 MHz光谱仪的灵敏度和色散增加 文书将对我们解决许多问题的能力产生重大影响， 问题增加的分散可能对以下方面最重要： 不能用15 N或13 C标记的系统， 必须使用1H NMR实验;此类系统的示例包括肽 蛋白质折叠中间体模型，蛋白质熔融球状态， 和DNA霍利迪连接。对于某些系统，如多个锌指 蛋白质，蛋白质-DNA复合物和RNA，同位素标记和 750 MHz频谱仪的高色散可能更大， 重要性仪器灵敏度的提高也将 对于仅限于集中的项目特别有利 1 mM或更低，这是由于溶解性或聚集的问题。