Upgrade of a Core Laboratory Macromolecular X-ray Diffraction System

项目摘要 这项共享仪器资助计划的目的是提高大分子X- 德克萨斯大学圣安东尼奥健康科学中心(UT Health)的射线结晶学 结晶学核心实验室(XRC)，位于内华达大学生物化学与结构生物学系 乔·R·朗和特蕾莎·洛扎诺·朗医学院。XRC有20年的历史，是一个机构 负责研究的副总裁办公室下的研究核心，以及药物发现和结构的一部分 NCI指定的梅斯癌症中心下的生物共享资源。它提供了结晶，X- 射线衍射数据收集和大分子结构测定服务为各种不同的 项目遍及该机构、其中心和圣安东尼奥地区。该工具的用户为 癌症生物学、DNA损伤和修复、先天免疫、传染性 疾病，药物设计等等。 XRC的当前资源包括一台MicroMax 007HF X射线发生器，驱动两个端口，具有 VariMax-HR和HF共焦光学元件瞄准两个安装在R轴上的HTC成像板探测器。007HF 旋转阳极发电机一直忠实地为XRC用户服务，并将继续运行和支持多年，以 来到制造商Rigaku(自1951年开始营业)。然而，成像板探测器已经被 被Rigaku逐步淘汰，从今年开始不再有资格获得部件支持。既要继续又要 加强XRC运作，我们建议将两个探测器和光学元件升级/更换为一个HyPix- 安装在万能Kappa测角仪和VariMax VHF共焦上的6000He光子计数探测器 具有连续可调发散狭缝组件的微聚焦光学元件。新的仪器将是 安装在现有的X射线发生器上，取代了这两个过时的成像板系统，还将 使用现有的牛津Cryostream 800冷却系统。成像版技术经历了两代 技术落后，被电荷耦合器件(CCD)探测器和现在的光子计数所取代 探测器。HyPix-6000HE的性能显著优于成像板技术，并优于CCD 光子计数探测器在消除误差同时具有高灵敏度和动态范围技术 由于读出噪声和暗电流。其他主要优势是数据的速度和数据量 由我们的成像板上的光子计数探测器获取。HyPix-6000HE可以获得完全 几分钟到几小时的时间范围内的数据集，而图像版需要几小时到几天的时间。总体而言， 要求升级到XRC将使调查人员能够快速、高质量、高分辨率地收集数据 在短时间内完成与人体健康相关的大分子的结构测定。

Project Summary The aim of this Shared Instrumentation Grant proposal is to enhance capabilities for macromolecular X- ray crystallography at the University of Texas Health Science Center at San Antonio (UT Health) X-ray Crystallography Core Laboratory (XRC), located in the Department of Biochemistry & Structural Biology within the Joe R. and Teresa Lozano Long School of Medicine. The XRC has a 20-year history and is an Institutional Research Core under the Office of the Vice President for Research and part of the Drug Discovery & Structural Biology Shared Resource under the NCI-designated Mays Cancer Center. It provides crystallization, X- ray diffraction data collection and macromolecular structure determination services for a wide variety of projects throughout the institution, its centers and the San Antonio area. Users for the instrumentation are investigators for topics such as cancer biology, DNA damage and repair, innate immunity, infectious disease, and drug design to name a few. Current resources in the XRC include a MicroMax 007HF X-ray generator driving two ports with VariMax-HR and HF confocal optics aimed at two mounted R-AXIS HTC imaging plate detectors. The 007HF rotating anode generator has faithfully served XRC users and will be functional and supported for years to come by the manufacturer Rigaku (in business since 1951). However, the imaging plate detectors have been phased out by Rigaku and are no longer eligible for parts support beginning this year. To both continue and enhance XRC operation, we propose to upgrade/replace the two detectors and optics with a single HyPix- 6000HE photon counting detector mounted on a Universal Kappa Goniometer and a VariMax VHF confocal microfocus optic with a continuously adjustable divergence slit assembly. The new instrumentation will be mounted on the existing X-ray generator, taking the place of both obsolete imaging plate systems, and will also use the existing Oxford Cryostream 800 cooling system. The imaging plate technology is two generations behind in technology, superseded by charge coupled device (CCD) detectors and now photon counting detectors. The HyPix-6000HE significantly outperforms imaging plate technology and improves upon CCD technology in that photon counting detectors have high sensitivity and dynamic range while eliminating error due to readout noise and dark current. Additional major advantages are the speed and volume of data that can be acquired by photon counting detectors over our imaging plates. The HyPix-6000HE can acquire full datasets in the timescale of minutes to hours while the image plates require hours to days. Overall, the requested upgrade to the XRC will benefit investigators with rapid, high quality, high resolution data collection to accomplish structure determinations of macromolecules relevant to human health in short order.