500 - MHZ REDOR NMR SPECTROMETER

500 - MHZ REDOR 核磁共振波谱仪

• 批准号: 6288313
• 负责人: JACOB SCHAEFER
• 金额: $ 50万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6288313
• 关键词: biomedical equipment development; biomedical equipment purchase; biotechnology; macromolecule; nuclear magnetic resonance spectroscopy; structural biology; technology /technique development

We are using new solid-state rotational-echo double resonance (REDOR) NMR experiments to characterize the structures of biologically and biomedically important macromolecular systems. These systems are not suited to analysis by diffraction or solution-state NMR methods. They are also beyond the current scope of the total-structure, solid-state NMR methods being developed by Opella, Griffin, Tycko, and others. We are developing a complementary NMR approach in which we combine available structural information about the systems or their subunits (from crystallography, solution-state NMR, electron diffraction, ab initio and homology-based calculations), with detailed REDOR information about restricted regions (interfaces, channels, binding sites) important to biological function. Currently there are 5 major users of the high-sensitivity, high-resolution 500-MHz REDOR spectrometer in the structural biology program at Washington University, and another 9 minor participants. The most severe limitation on progress in implementing our REDOR program is the available machine time. There is sufficient demand for machine time by the major users to occupy completely the present spectrometer for more than 18 months with existing samples and planned experiments. The average wait for access is 6 months. Two of the minor users have been waiting for a year for access and need time to conduct feasibility tests and collect preliminary data. Our solution to this dilemma is to create a new solid-state NMR laboratory with the help of an NIH shared-instrumentation grant and substantial support from Washington University. This facility will contain three 500-MHz REDOR spectrometers: the present machine and two clones. Adding two new spectrometers is first, cost effective (because of attractive prices for buying in bulk), and second, necessary to meet scheduling demands of both major and minor users. NIH funds will be used for one of the new spectrometers, and Washington University funds for the other. Although there is no suitable REDOR spectrometer sold by a single vendor, all of the components for the two new spectrometers are commercially available with the exception of the special REDOR probes. For these, we propose to build in-house, two 6- frequency probes with our proven (and patented), high-performance transmission-line probe technology. These probes will differ in the type of magic-angle spinning rotor employed. Thus, rotor volume and spinning speed can be tailored to the demands of specific experiments for enhanced sensitivity. We anticipate that this combination of new spectrometers and probes will multiply the productivity of our structural biology solid-state NMR laboratory by a factor of four and thereby eliminate the need for a queue for either major or minor users.

我们正在使用新的固态旋转回波双共振 (REDOR) NMR 实验来表征生物学和生物医学上重要的大分子系统的结构。这些系统不适合通过衍射或溶液态核磁共振方法进行分析。它们也超出了 Opella、Griffin、Tycko 等公司目前正在开发的全结构固态 NMR 方法的范围。我们正在开发一种互补的 NMR 方法，其中我们将有关系统或其亚基的可用结构信息（来自晶体学、溶液态 NMR、电子衍射、从头计算和基于同源性的计算）与对生物功能重要的限制区域（界面、通道、结合位点）的详细 REDOR 信息结合起来。目前，华盛顿大学结构生物学项目中高灵敏度、高分辨率500 MHz REDOR光谱仪有5个主要用户，另有9个次要参与者。实施 REDOR 计划的最严重限制是可用的机器时间。主要用户对机器时间的需求足以使现有的光谱仪完全占用现有样品和计划的实验超过 18 个月。平均等待访问时间为 6 个月。其中两个未成年用户已经等待了一年的访问，需要时间进行可行性测试并收集初步数据。我们解决这一困境的方法是在 NIH 共享仪器拨款和华盛顿大学的大力支持下创建一个新的固态 NMR 实验室。该设施将包含三台 500 MHz REDOR 光谱仪：现有机器和两台克隆机。添加两台新的光谱仪首先是具有成本效益（因为批量购买的价格具有吸引力），其次是满足主要和次要用户的调度需求所必需的。美国国立卫生研究院的资金将用于其中一台新的光谱仪，华盛顿大学的资金将用于另一台。尽管没有单一供应商销售合适的 REDOR 光谱仪，但除了特殊的 REDOR 探头外，这两种新型光谱仪的所有组件都可以在市场上买到。为此，我们建议使用我们经过验证（且已获得专利）的高性能传输线探头技术构建两个内部 6 频率探头。这些探头的不同之处在于所采用的魔角旋转转子的类型。因此，转子体积和旋转速度可以根据特定实验的要求进行定制，以提高灵敏度。我们预计，新型光谱仪和探头的组合将使我们结构生物学固态 NMR 实验室的生产力提高四倍，从而消除主要或次要用户排队的需要。