Laboratory of Biomolecular Structure and Function

项目总结 (生物分子结构与功能实验室) OUHSC X射线核心设备是在1998年启动的，NIH向保罗·魏格尔博士提供了创意拨款，他曾担任 生物化学和分子生物学系。该设施最近更名为 生物分子结构与功能(LBSF)LBSF提供专业知识和评估工具，以 个别科布雷项目研究对结晶学和小角X射线至关重要的各种技术 蛋白质和核酸的散射研究(SAXS)结晶学技术包括筛选 室温衍射性晶体，低温晶体保护剂筛选，晶体低温保护 高压下的衍射数据采集、数据处理、空间群归属、结构 确定、结构细化、结构分析、结构解释和图形准备。这个 与SAXS相关的方法包括动态光散射以检查聚集体和测量多分散性 在将样品送往国家设施进行SAXS数据收集之前。在科布雷的第一阶段，设施 升级后配备了新的Rigaku MicroMax 007 X射线发生器，平板怀亚特平板读数动态灯 用于高通量研究的散射仪和用于制作晶体图像的徕卡显微镜。这个 新的X射线发生器提供了四倍强度更高、直径更小的X射线束。这些功能至关重要 用于筛选Cobre研究项目中常见的小而弱的衍射晶体。这个 俄克拉荷马医学研究基金会捐赠了一台Mar345图像板X射线解析器，一台Varimax HF光学设备 用于将惰性气体引入质子腔的系统、牛津冷冻系统和MSC Xe-Site装置 用于相变实验的晶体中。对Rigaku X射线系统进行了改进，增加了第二个工作台 将Mar345系统添加到X射线发生器的第二个端口。RaxisIV系统是 封闭有屏蔽，使用户能够在收集数据时将晶体安装在MAR系统上 在Rigaku系统上。此设置使两个组能够同时收集数据，或者使一个组能够 同时采集两个晶体的衍射数据。新的X射线发生器已被用于筛查 蛋白质和RNA晶体的衍射质量和发展低温条件。读盘式DLS 已使用设备来筛选那些促进蛋白质稳定性的缓冲液以用于蛋白质存储和 结晶试验。DLS还被用于筛选SAXS样本，以确定是否存在聚集体 运往国家设施的SAXS光束线。在第一阶段，十个小组参与了SAXS研究。在.期间 第二阶段，将增加一个综合分子模型组，以方便密集的计算工作 作为候选药物分子的虚拟筛选、分子动力学模拟、分子置换 以及具有系综结构的SAXS数据的分子模拟。这个单位将是 每天由OMRF的Tim Mather博士管理，他也是OUHSC的兼职成员 生物化学与分子生物学系。此外，一款Art Robbins结晶机器人 脂质立方相中完整膜蛋白的结晶将添加到LBSF中，以促进 OUHSC校园膜蛋白的结晶学研究。MCL On中的结晶机器人 俄亥俄大学诺曼分校将继续用于可溶性蛋白质的研究。MCL还将用于高- 用Platmate和Pilatus探测器筛选晶体的产量。OUHSC一直在为 西蒙·特尔兹安博士是一名工作人员，他于2015年受聘管理LBSF的日常运作。 他维护仪器，安排用户，并提供结晶学方面的培训。他还安装了 大多数新乐器。2015年，LBSF成为由 研究副总裁(VPR)。该套件的管理核心负责开具发票并管理 LBSF的独立账户。VPR为LBSF设立了一个监督委员会。这个委员会是 由科布雷副主任担任主席。LBSF主任布莱恩·穆尔斯博士负责这方面的工作 委员会作为临时成员，在研讨会上推广LBSF，监督实验室经理，写道 负责设备补助申请，并为VPR、EAC和NIH撰写进度报告。他一直是 自2008年以来一直担任该设施的主任。Mooers和Terzyan博士拥有超过45年的联合经验 大分子结晶学将帮助项目研究人员在他们提出的 第二阶段的结构研究。

PROJECT SUMMARY (Laboratory for Biomolecular Structure and Function) The OUHSC X-ray Core Facility was initiated in 1998 with a NIH IDeA grant to Dr. Paul Weigel, former chair of the Department of Biochemistry and Molecular Biology. The facility was recently renamed the Laboratory of Biomolecular Structure and Function (LBSF). The LBSF provides expertise and assess to instruments to the individual COBRE projects on a variety of techniques critical to crystallographic and small angle X-ray scattering studies (SAXS) of proteins and nucleic acids. The crystallographic techniques include screening crystals for diffraction quality at room temperature, crystal cryo-protectant screening, crystal cryprotection under high pressure, diffraction data collection, diffraction data processing, space group assignment, structure determination, structure refinement, structure analysis, structure interpretatoin, and figure preparation. The SAXS-related methods include dynamic light scattering to check for aggregates and to measure polydispersity prior to sending the samples for SAXS data colleciton at national facilities. In Phase I of the COBRE, the facility was upgraded with a new Rigaku MicroMax 007 X-ray generator, a plate Wyatt plate-reading dynamic light scattering instrument for high throughput studies, and a Leica microscope for making images of crystals. The new X-ray generator gives a four-fold more intense X-ray beam of smaller diameter. These features are critical for the screening the small, weakly diffracting crystals common in the COBRE research projects. The Oklahoma Medical Research Foundation donated a Mar345 image plate X-ray dectector, a Varimax HF optic system, an Oxford cryosystem, and a MSC Xe-Siter apparatus for introducing noble gases into protien cavities in crystals for phasing experiments. The Rigaku X-ray system was modifed by adding a second bench to enable the addition of the Mar345 system to the second port of the X-ray generator. The RaxisIV system was enclosed with shielding that enables users to mount crystals on the Mar system while data are being collected on the Rigaku system. This setup has enabled two groups to collect data simultaneously or one group to collect diffraction data from two crystals at the same time. The new X-ray generator has been used to screen protein and RNA crystals for diffraction quality and to develop cryo-conditions. The plate-reading DLS apparatus has been used to screen buffers for those that promote protein stability for protein storage and crystallization trials. DLS has also been used to screen SAXS samples for the presence of aggregates prior to shipment to SAXS beam lines at national facilities. In Phase I, ten groups participated in SAXS studies. During Phase II, an Integrative Molecular Modeling Unit will be added to faciltate intenstive computational work such as virutal screening for candidate drug molecules, molecular dynamics simulations, molecular replacement with phenix-rosetta, and molecular modeling of SAXS data with ensenbles of structures. This unit will be managed on a daily basis by Dr. Tim Mather from OMRF who is also an adjunct member of the OUHSC Department of Biochemistry and Molecualr Biology. In addtion, an Art Robbins crystallization robot for the crystallizaton of integral membrane proteins in the lipidic cubic phase will be added to the LBSF to facilitate crystallograpic studies of membrane proteins on the OUHSC campus. The crystallization robots in the MCL on the OU-Norman campus will continue to be used for soluble proteins. The MCL will also be used for high- troughput screening of crystals with the Plate-Mate and Pilatus detector. OUHSC has been providing salary for a staff sceintist, Dr. Simon Terzyan, who was hired in 2015 to manage the day-to-day operatons of the LBSF. He maintains the instruments, schedules users, and provides training in crystallography. He has also installed most of the new instruments. In 2015, the LBSF became a core lab in the suite of core labs supported by the Vice President of Research (VPR). The administrative core of this suite issues invoices and manages the independent account of the LBSF. The VPR setup an oversight committee for the LBSF. This committee is chaired by the COBRE associate director. The Director of the LBSF, Dr. Blaine Mooers, serves on this committee as an ad hoc member, promotes the LBSF in seminars, oversees the Lab Manager, writes equipment grant applications, and writes progress reports for the VPR, EAC, and NIH. He had been the director of this facility since 2008. Drs Mooers and Terzyan have over 45 years of combined experience in macromolecular crystallography that will help the Project Investigators achieve success in their proposed structural studies during Phase II.