A NEW HYBRID DIFFRACTOMETER FOR MACROMOLECULAR CRYSTALLOGRAPHY AND SAXS AT TJU

天津大学用于高分子晶体学和 SAXS 的新型混合衍射仪

• 批准号: 8639253
• 负责人: Gino Cingolani
• 金额: $ 60万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8639253
• 关键词: Anodes; Area; Basic Science; Biomedical Research; Cancer Biology; Cancer Center; Core Facility; Cost Sharing; Crystallography; Data; Data Collection; Development; Doctor of Philosophy; Equipment; Failure; Fees; Funding; Goals; Housing; Hybrids; Image; Institution; Left; Light; Maintenance; Minor; Mission; Molecular Biology; Research; Research Project Grants; Resource Sharing; Roentgen Rays; Services; Solutions; Source; Spottings; Staging; Synchrotrons; System; Techniques; Testing; Tube; United States National Institutes of Health; Universities; Wages; Work; X-Ray Crystallography; aged; anticancer research; beamline; clinical application; cryogenics; detector; improved; instrumentation; medical schools; programs; screening; seal; structural biology

DESCRIPTION (provided by applicant): The X-ray Crystallography and Macromolecular Characterization Shared Resource Facility at the Kimmel Cancer Center (KCC), dedicated to supporting cutting edge structural and molecular biology research applied to cancer biology, is one of the 11 NCI-supported Core Facilities at Thomas Jefferson University (TJU). Because of its potential to enhance existing NIH-funded research programs and further develop research at TJU research centers, X-ray crystallography is one of the techniques identified by Dr. Tykocinski, our Dean of Jefferson Medical College, for strategic development of our research mission. Our existing two X-ray generators are between 11 and 22 years old, limiting our current ability to carry out cutting edge structural work. The older diffractometer (installed in Oct 1991) was decommissioned to users in Sept 2010 due to constant failure in the rotating anode. The younger unit, a sealed X-ray tube operational since Feb 2002, is also prone to frequent service failures, costly maintenance and provides a very weak X-ray beam. Therefore, as part of a cost-sharing plan to establish a state-of-the-art center for structural biology at TJU, we request funds to replace both aged diffractometers with a Rigaku MicroMax-007HF microfocus rotating anode X-ray generator dual equipped for data collection from macromolecular crystals and Small Angle X-Ray Scattering (SAXS). We have identified a group of 9 major core users and 2 minor users from both inside TJU and in nearby institutions who eagerly support this proposal. If funded, TJU will leverage this support with ~$280,000 in parts derived from our current Rigaku diffractometer, and funds supporting 30% of a Ph.D.-level facility manager, Dr. Anshul Bhardwaj. The KCC will also infuse four years of additional funds to cover the remaining 70% of Dr. Bhardwaj salary and equipment maintenance fees. In the planning stages, our preliminary tests indicated that the Rigaku MicroMax-007HF is ideally suited for our needs. This system allows us to re-utilize perfectly functional parts of our current generator (i.e. R-AXIS IV++ imaging plate area detector, XStream 2000 cryogenic system, etc.) in the left port, while the right port will host the new BioSAXS-1000 unit. With a unique 70 um circular focal spot, the MicroMax-007HF is perfectly suited for rapid in house screening of small crystals. Preliminary tests have also shown that the Rigaku BioSAXS-1000 unit provided SAXS data of comparable quality as beamline X9 at the National Synchrotron Light Source (NSLS). Additionally, the routine maintenance needs of the proposed Rigaku system are lower than that of our combined existing equipment. By increasing X-ray beam intensity and expanding our capabilities for in solution SAXS analysis, the proposed instrumentation will significantly enhance the competitiveness of current NIH-funded research projects and expand the body of users to our NCI-funded facility, with the ultimate goal of improving the quality of our biomedical research, and strengthening the liaison between basic research, cancer research and its clinical applications.

描述（由申请人提供）：Kimmel 癌症中心 (KCC) 的 X 射线晶体学和大分子表征共享资源设施致力于支持应用于癌症生物学的尖端结构和分子生物学研究，是 NCI 支持的 11 个项目之一托马斯杰斐逊大学 (TJU) 的核心设施。由于 X 射线晶体学具有增强现有 NIH 资助的研究项目并进一步发展 TJU 研究中心研究的潜力，因此它是杰斐逊医学院院长 Tykocinski 博士确定的技术之一，用于战略发展我们的研究任务。我们现有的两台 X 射线发生器的使用年限为 11 至 22 年，限制了我们目前开展尖端结构工作的能力。较旧的衍射仪（1991 年 10 月安装） 由于旋转阳极不断出现故障，该设备于 2010 年 9 月停止向用户使用。较新的装置是自 2002 年 2 月开始运行的密封 X 射线管，也容易出现频繁的服务故障、维护成本高昂，并且提供的 X 射线束非常弱。因此，作为在 TJU 建立最先进的结构生物学中心的成本分摊计划的一部分，我们请求资金 用 Rigaku MicroMax-007HF 微焦点旋转阳极 X 射线发生器替换两台老化的衍射仪，该发生器双重配备用于从大分子晶体和小角度 X 射线散射 (SAXS) 收集数据。我们从 TJU 内部和附近机构中确定了 9 位主要核心用户和 2 位次要用户，他们热切支持该提案。如果获得资助，TJU 将利用这一支持，提供约 280,000 美元的零件，这些零件来自我们当前的 Rigaku 衍射仪，并为博士级设施经理 Anshul Bhardwaj 博士提供 30% 的资金支持。 KCC 还将注入四年的额外资金，用于支付 Bhardwaj 博士剩余 70% 的工资和设备维护费。在规划阶段，我们的初步测试表明 Rigaku MicroMax-007HF 非常适合我们的需求。该系统使我们能够在左侧端口重新利用当前发生器的完美功能部件（即 R-AXIS IV++ 成像板区域探测器、XStream 2000 低温系统等），而右侧端口将托管新的 BioSAXS-1000单元。 MicroMax-007HF 具有独特的 70 um 圆形焦点，非常适合小晶体的快速内部筛选。初步测试还表明，Rigaku BioSAXS-1000 装置提供的 SAXS 数据质量与国家同步加速器光源 (NSLS) 的光束线 X9 相当。此外，拟议的 Rigaku 系统的日常维护需求低于我们现有设备的组合。通过增加 X 射线束强度并扩展我们的解决方案 SAXS 分析能力，拟议的仪器将显着提高当前 NIH 资助的研究项目的竞争力，并将用户群体扩大到我们 NCI 资助的设施，最终目标提高生物医学研究质量，加强基础研究、癌症研究与临床应用的衔接。

项目成果

Structure of the pre-mRNA leakage 39-kDa protein reveals a single domain of integrated zf-C3HC and Rsm1 modules.

前体 mRNA 泄漏 39-kDa 蛋白的结构揭示了集成 zf-C3HC 和 Rsm1 模块的单个结构域。

• 发表时间: 2022-10-21
• 影响因子: 4.6
• 作者: Hashimoto, Hideharu;Ramirez, Daniel H;Lautier, Ophélie;Pawlak, Natalie;Blobel, Günter;Palancade, Benoît;Debler, Erik W
• 通讯作者: Debler, Erik W

The effects of molecular crowding and CpG hypermethylation on DNA G-quadruplexes formed by the C9orf72 nucleotide repeat expansion.

分子拥挤和 CpG 超甲基化对 C9orf72 核苷酸重复扩展形成的 DNA G 四链体的影响。

• 发表时间: 2021-12-01
• 影响因子: 4.6
• 作者: Ozcan, Kadir A;Ghaffari, Layla T;Haeusler, Aaron R
• 通讯作者: Haeusler, Aaron R

Three-dimensional structure of human cyclooxygenase (hCOX)-1.

人环氧合酶（hCOX）-1的三维结构。

• 发表时间: 2021
• 影响因子: 4.6
• 作者: Miciaccia, Morena;Belviso, Benny Danilo;Iaselli, Mariaclara;Cingolani, Gino;Ferorelli, Savina;Cappellari, Marianna;Loguercio Polosa, Paola;Perrone, Maria Grazia;Caliandro, Rocco;Scilimati, Antonio
• 通讯作者: Scilimati, Antonio