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声双共振（REDOR）核磁共振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。这些体系不适合用衍射或溶液态核磁共振方法进行分析。它们也超出了目前由Opella、Griffin、Tycko等人开发的全结构固态核磁共振方法的范围。我们正在开发一种互补的核磁共振方法，在该方法中，我们将有关系统或其亚基的可用结构信息（来自晶体学，溶液态核磁共振，电子衍射，从头算和基于同源的计算）与有关生物功能重要的受限区域（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的高灵敏度、高分辨率500-MHz REDOR光谱仪有5个主要用户，另外还有9个次要参与者。在实施我们的REDOR程序的过程中，最严重的限制是可用的机器时间。主要用户对机器时间有足够的需求，以现有样品和计划的实验完全占用现有光谱仪18个月以上。平均等待访问时间为6个月。其中两个小用户已经等待了一年，需要时间进行可行性测试和收集初步数据。我们解决这一困境的办法是在NIH共享仪器的资助和华盛顿大学的大力支持下，创建一个新的固态核磁共振实验室。该设施将包含三个500-MHz的REDOR光谱仪：目前的机器和两个克隆。增加两台新的光谱仪首先具有成本效益（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。其中一台新的光谱仪将由美国国立卫生研究院资助，另一台由华盛顿大学资助。虽然没有合适的REDOR光谱仪由单一供应商销售，但除了特殊的REDOR探头外，这两个新光谱仪的所有组件都是市售的。为此，我们建议使用我们成熟的（和专利的）高性能在线传输探针技术构建内部两个6频率探头。这些探头在使用的魔角旋转转子类型上有所不同。因此，转子体积和旋转速度可以根据特定实验的要求进行调整，以提高灵敏度。我们预计，这种新的光谱仪和探针的组合将使我们的结构生物学固态核磁共振实验室的生产力增加四倍，从而消除了为主要或次要用户排队的需要。