Upgrade of In-house X-ray Diffraction Equipment

内部X射线衍射设备升级

• 批准号: 7387985
• 负责人: DANIEL J LEAHY
• 金额: $ 41.9万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7387985
• 关键词: Biophysics; Caliber; Chemicals; Collaborations; Crystallography; Data; Data Collection; Drug Design; Equipment; Funding; Gene Expression; Gene Expression Regulation; Housing; Image; Institution; Methods; Optics; Oxidoreductase; Pharmacology; Publishing; Range; Reaction; Research; Roentgen Rays; Signal Transduction; Stream; Structural Biologist; Structure; System; Testing; Time; Transduction Gene; Universities; X ray diffraction analysis; X-Ray Diffraction; base; biophysical chemistry; detector; enzyme mechanism; medical schools; nucleic acid metabolism; research study; size; tool

DESCRIPTION (provided by applicant): Three groups in the Department of Biophysics & Biophysical Chemistry at the Johns Hopkins University School of Medicine (JHUSOM) use X-ray diffraction as a primary research tool (Amzel, mechanism of oxidoreductases, phosphoryl transfer reactions, and macromolecular recognition; Leahy, signal transduction; Wolberger, regulation of gene expression), and each of these groups collaborates with multiple groups at Hopkins as well as outside institutions. In the past several years two groups in the Department of Pharmacology at Hopkins (Cole, chemical approaches to signal transduction and gene expression; Stivers, nucleic acid metabolism) have started to use diffraction methods as a significant component of their research. These efforts began as collaborations with structural biologists at Hopkins and elsewhere but are expanding so that X-ray structure determinations are becoming a component of their independent research efforts. The current X-ray diffraction facilities at JHUSOM are heavily oversubscribed, and we propose to take advantage of recent and impressive advances in in-house X-ray generators to increase the capacity and capabilities of this facility. Specifically, we propose to acquire a Rigaku 007HF high-flux X-ray generator equipped with two sets of VariMax HF optics, one for each port on the 007HF generator. We also propose to acquire an R-axis IV++ Image Plate detector to replace an outmoded and unsupported R-axis II detector. An existing R-axis IV detector and two X-stream cryo-cooling systems will be transferred to the new generator along with the R-axis IV++ to create two high-flux X-ray stations for macromolecular crystallography. Based on test crystal comparisons and published specifications, we expect the new generator and optics to result in a 10-fold decrease in average data collection times and a substantial increase in project throughput. Owing to the decreased beam size produced by the new generator (70 5M diameter) vs. the existing equipment (300 5M diameter) and increased X-ray beam intensity, the new setup will be especially advantageous for data collection from small crystals, which make up the majority of current experiments. By increasing throughput and expanding the range of projects for which usable diffraction data can be obtained, the upgraded facility will significantly enhance a broad range of NIH-funded, health-related projects with implications for signaling, gene expression, enzyme mechanism, and drug design.

描述（申请人提供）：约翰霍普金斯大学医学院生物物理与生物物理化学系（JHUSOM）的三个小组使用X射线衍射作为主要研究工具（Amzel，氧化还原酶、磷酰基转移反应和大分子识别的机制; Leahy，信号转导; Wolberger，基因表达调控），这些小组中的每一个都与霍普金斯的多个小组以及外部机构合作。在过去的几年里，霍普金斯药理学系的两个研究小组（科尔，信号转导和基因表达的化学方法;斯提弗斯，核酸代谢）已经开始使用衍射方法作为他们研究的重要组成部分。这些努力开始于与霍普金斯和其他地方的结构生物学家的合作，但正在扩大，使X射线结构测定成为他们独立研究工作的一部分。JHUSOM目前的X射线衍射设施严重超额认购，我们建议利用内部X射线发生器的最新和令人印象深刻的进步来提高该设施的容量和能力。具体来说，我们建议购买一台配备两套VariMax HF光学器件的理学007 HF高通量X射线发生器，007 HF发生器上的每个端口一套。我们还建议购买R轴IV++图像板探测器，以取代过时且不受支持的R轴II探测器。现有的R轴IV探测器和两个X流低温冷却系统将与R轴IV++一起转移到新的发生器沿着，以创建两个用于大分子晶体学的高通量X射线站。根据测试晶体的比较和已发布的规格，我们预计新的发生器和光学器件将使平均数据收集时间减少10倍，并大幅提高项目吞吐量。由于新发生器（70 5 M直径）与现有设备（300 5 M直径）相比产生的光束尺寸减小，并且X射线光束强度增加，新设置将特别有利于从小晶体收集数据，这构成了当前实验的大部分。通过增加吞吐量和扩大可以获得可用衍射数据的项目范围，升级后的设施将显着增强NIH资助的广泛的健康相关项目，这些项目对信号传导，基因表达，酶机制和药物设计具有影响。