X-ray Source Upgrade for Macromolecular Crystallography

高分子晶体学 X 射线源升级

• 批准号: 8051488
• 负责人: David B McKay
• 金额: $ 11.22万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2012-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8051488
• 关键词: Anodes; Biological Process; Catalytic RNA; Colorado; Complex; Crystallization; Crystallography; DNA; Data; Data Collection; Data Set; Funding; Gifts; Housing; Humpback Dolphins; Integral Membrane Protein; Minor; Molecular Chaperones; Phase; Proteins; RNA; Resolution; Roentgen Rays; Screening procedure; Source; Staging; Synchrotrons; System; Telomerase; United States National Institutes of Health; Universities; protein complex

DESCRIPTION (provided by applicant): The macromolecular x-ray diffraction facility of the University of Colorado, Boulder proposes to upgrade an X-ray source: specifically, a Rigaku RUH2R rotating anode X-ray generator with Osmic "blue" confocal multilayer mirrors will be upgraded to a Rigaku MM007 generator with Varimax HR (="high resolution") confocal mirrors. We are receiving a used MM007 as a gift, and request funds only for the confocal mirrors. The x-ray facility is currently used by three major and seven minor NIH-supported users. Crystallographic projects currently underway include: large catalytic RNAs (Cech and Pace) and riboswitches (Batey), RNA- protein and DNA-protein complexes such as those found in the telomerase system (Cech and Wuttke) and specific recognition complexes (McKay), molecular chaperone and integral membrane proteins (Sousa), and several RNAs and proteins involved in specific biological processes (Ahn, Copley, Pardi, Falke). Many of these projects pose significant challenges in their early stages; typically, small preliminary crystals from a large number of constructs and/or crystallization conditions must be screened for diffraction to determine whether they are worth pursuing. Additional challenges often present themselves during screens for cryoprotectants or suitable phasing derivatives. Finally, in many cases, complete datasets are collected in- house, either prior to or as an alternative to synchrotron data collection. The requested upgrade will enhance the brightness of our X-ray source severalfold (approximately sixfold according to Rigaku's data), thereby enhancing the crystal screening and data collection capability of our in-house facilities. PUBLIC HEALTH RELEVANCE: The three-dimensional structures of biologically significant macromolecules provide the foundation for modern molecular medicine and rational drug design. Structural studies at the University of Colorado, Boulder encompass ribozymes (catalytic RNAs), telomerase, membrane protein structure and folding, and bacterial drug resistance, to name a few areas. The shared X-ray crystallography facility, for which instrumentation is requested, provides resources (both instrumental and intellectual) to a broad spectrum of users who pursue structural studies on the campus.

描述（由申请人提供）： 科罗拉多大学博尔德分校的高分子 X 射线衍射设施提议升级 X 射线源：具体来说，配备 Osmic“蓝色”共焦多层镜的 Rigaku RUH2R 旋转阳极 X 射线发生器将升级为配备 Varimax HR（“高分辨率”）共焦镜的 Rigaku MM007 发生器。我们收到了二手 MM007 作为礼物，并且仅要求为共焦镜提供资金。该 X 射线设备目前由 NIH 支持的 3 个主要用户和 7 个次要用户使用。目前正在进行的晶体学项目包括：大型催化RNA（Cech和Pace）和核糖开关（Batey）、RNA-蛋白质和DNA-蛋白质复合物，例如端粒酶系统中发现的RNA-蛋白质和DNA-蛋白质复合物（Cech和Wuttke）和特异性识别复合物（McKay）、分子伴侣和整合膜蛋白（Sousa），以及参与特定生物过程的几种RNA和蛋白质 （安、科普利、帕迪、法尔克）。其中许多项目在早期阶段都面临着重大挑战；通常，必须对来自大量构建体和/或结晶条件的小型初步晶体进行衍射筛选，以确定它们是否值得追求。在筛选冷冻保护剂或合适的定相衍生物时经常会遇到额外的挑战。最后，在许多情况下，完整的数据集是在内部收集的，无论是在同步加速器数据收集之前还是作为同步加速器数据收集的替代方案。所要求的升级将使我们的X射线源的亮度提高数倍（根据理学的数据约为六倍），从而增强我们内部设施的晶体筛选和数据收集能力。 公共卫生相关性：具有生物学意义的大分子的三维结构为现代分子医学和合理的药物设计奠定了基础。科罗拉多大学博尔德分校的结构研究包括核酶（催化 RNA）、端粒酶、膜蛋白结构和折叠以及细菌耐药性等领域。需要配备仪器的共享 X 射线晶体学设施为在校园内进行结构研究的广大用户提供资源（包括仪器资源和知识资源）。