UPGRADE OF IN-HOUSE X-RAY DIFFRACTION EQUIPMENT: STRUCTURAL BIOLOGY

该子项目是利用由NIH/NCRR资助的共享仪器补助金提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是资助机构，不一定是研究者的机构。描述（由申请人提供）：“在洛克菲勒大学，有三个相对较大、非常成功和富有成效的结构生物学小组，他们使用x射线晶体学作为主要工具（Darst，细菌转录结构生物学；MacKinnon，离子通道结构生物学；Stebbins，细菌致病机理结构生物学）。在过去5年左右的时间里，洛克菲勒的另外三个小组开发了重要的x射线晶体学项目（Blobel， mRNA核输出；Rice，病毒复制；Sakmar， G蛋白偶联信号）。这些努力开始于与洛克菲勒的结构生物学家的正式或非正式的合作，但在每个案例中，成功都导致了这些努力的扩展，例如这些团体支持x射线晶体学计划，就像结构生物学家一样庞大和活跃。这种情况导致洛克菲勒大学需要对内部x射线设备进行重大升级。具体来说，该共享仪器补助金提出了以下升级：1)一个R-Axis IV++区域检测器，以取代一个使用了12年的R-Axis II区域检测器，该区域检测器将不再被MSC支持；2)逆<)>括号加到x射线发生器的两个端口；3) 2套VariMax Cu HR共聚焦Max-Flux光学系统，Rigaku/MSC提供的最先进的x射线光学系统，取代使用了12年的MSC/Yale镜；4)两个x流低温系统，以取代过时的、已有15年历史的MSC低温系统。这些升级将大大提高机器筛选膜蛋白和大分子复合物弱衍射晶体的使用，提高数据质量和通量，并大大促进其用于筛选和制备用于同步加速器的冷冻保存晶体，这是nih支持的许多项目的主要用途。

This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): " At The Rockefeller University, there are three relatively large, highly successful and productive structural biology groups that use X-ray crystallography as a major tool (Darst, structural biology of bacterial transcription; MacKinnon, structural biology of ion channels; Stebbins, structural biology of bacterial pathogenesis). In the last 5 years or so, three additional groups at Rockefeller have developed significant X-ray crystallography programs (Blobel, mRNA nuclear export; Rice, viral replication; Sakmar, G protein-coupled signalling). These efforts began as formal or informal collaborations with the structural biologists at Rockefeller, but in each case success has led to the expansion of these efforts, such that these groups support X-ray crystallography programs as large and active as the structural biologists. This situation has led to a need for significant upgrades of the in-house X-ray equipment at The Rockefeller University. Specifically, this Shared Instrumentation Grant proposes the following upgrades: 1) An R-Axis IV++ area detector to replace a 12-year-old R-Axis II area detector that will no longer be supported by MSC; 2) Inverse <)> brackets to add to two ports of the X-ray generator; 3) Two VariMax Cu HR confocal Max-Flux optical systems, the most advanced X-ray optics system available from Rigaku/MSC, to replace 12-year-old MSC/Yale mirrors; 4) Two X-stream cryogenic systems to replace obsolete, 15-year-old MSC cryosystems that are out of repair. These upgrades will significantly enhance the use of the machine for screening weakly diffracting crystals of membrane proteins and large macromolecular complexes, enhance data quality and throughput, as well as greatly facilitate its use for the screening and preparation of cryo-preserved crystals for synchrotron trips, which is the main use of the machines for a number of NIH-supported projects.