GM/CA@APS: A Macromolecular Crystallography Resource

GM/CA@APS：高分子晶体学资源

• 批准号: 10251998
• 负责人: ROBERT F. FISCHETTI
• 金额: $ 422.77万
• 依托单位: ARGONNE NATIONAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10251998
• 关键词: 3-Dimensional; Address; Aging; Attention; Binding; Biochemistry; Caliber; Cancer Etiology; Capital; Cells; Collimator; Communities; Community Outreach; Computer software; Coupled; Crystallization; Crystallography; DNA; Data; Data Analyses; Data Collection; Data Storage and Retrieval; Deposition; Detection; Devices; Educational workshop; Ensure; Environment; Equipment; Failure; Feedback; Floor; Fluorescence; Funding; Goals; Immune system; Individual; Infection; Intuition; Knowledge; Laboratories; Macromolecular Complexes; Maintenance; Measures; Membrane; Methodology; Methods; Michigan; Modernization; Mutation; National Institute of General Medical Sciences; Nature; Noise; Optics; Organism; Paper; Peer Review; Performance; Periodicity; Pharmaceutical Preparations; Photons; Positioning Attribute; Preparation; Proteins; Protocols documentation; Publishing; Resolution; Resources; Robotics; Roentgen Rays; Running; Sampling; Scanning; Schedule; Schools; Science; Site; Source; Speed; Stimulus; Structural Protein; Structure; System; Technology; Time; Training; Universities; X-Ray Crystallography; base; beamline; cluster computing; computerized data processing; computing resources; crystallinity; data analysis pipeline; detector; experience; experimental study; graphical user interface; improved; instrument; instrumentation; operation; outreach; pathogen; programs; protein complex; protein structure; receptor; repaired; screening; structural biology; success; three dimensional structure; tool; tumor progression; user-friendly

The primary goal of the GM/CA@APS Resource is to provide user access to cutting-edge macromolecular crystallography beamlines to determine 3-D atomic structures of forefront problems in structural biology. The Resource was founded in 1999 and has been in operation as a national user facility since 2004, providing a full array of cutting-edge technologies for macromolecular crystallography in Sector 23 at the Advanced Photon Source (APS), Argonne National Laboratory. GM/CA is led by Robert Fischetti (Argonne National Laboratory, contact PI) and Janet Smith (University of Michigan, PI) with an effective administrative structure to operate, maintain, and upgrade the beamlines, and run a high-impact user program. The GM/CA macromolecular crystallography core provides a modern, powerful environment with sophisticated tools for sample screening, interrogation and data collection in fixed-energy, SAD and MAD modes through operation of two insertion-device X-ray beamlines, 23ID-B and 23ID-D, from canted-undulator sources. The beamlines are independently tunable over a wide energy range and are built to deliver a brilliant X-ray beam with exceptionally stable position and intensity. The exceptional stability makes GM/CA an ideal facility for anomalous scattering, and users can quickly (few minutes) change energy at will within the 5.0-20 keV range (λ = 2.48 Å - 0.62 Å). The stable beam enables a powerful microcrystallography capability. Small beams for microcrystallography are created by a quad mini-beam collimator with user-selectable beam sizes of 5-, 10- and 20-micron diameter in addition to the full beam. Users can rapidly (within seconds) change beam size for any sample at any time without beam realignment. The beamlines are equipped with fast, low-noise pixel-array detectors, an Eiger 16M on 23ID-B and a Pilatus3 6M on 23ID-D. Cryo-protected samples are robotically mounted from a 288-sample Dewar and can be visualized and manipulated in the beam with micron precision. Users control the beamlines and samples with the intuitive and biologist-friendly JBluIce graphical user interface. JBluIce includes GUI-based functions for beamline control, sample centering, sample screening, data collection, X-ray fluorescence energy scan, raster scan of the sample, and data analysis. The powerful raster feature is used to scan fields of micro-crystals or to find the best diffracting region of a larger crystal. Information from raster scans can be shared with the data collection routine for a variety of data collection options. The data analysis routines provide unit cell and resolution estimates to formulate a strategy for data collection, which is automatically coupled to three data processing pipelines, providing users with feedback in near realtime. During the 2016-2018 period, a total of 857 unique individuals from 157 unique groups used GM/CA beamlines, and annually interrogated 35,000 samples, deposited 225 structures in the PDB, and published 130 papers (12% in Nature, Science, and Cell). Serial crystallography methodologies are implemented and integrated into the JBluIce automated data collection and processing pipelines for both viscous injector and fixed-target sample delivery technologies. A computing cluster and storage array network provides high-speed data storage, retrieval, backup, and realtime data processing. Users receive training from an expert staff, and can collect data on-site or remotely. The staff also contribute to a variety of community outreach activities including the annual CCP4/APS Crystallography School. A biochemistry lab provides essential capabilities for sample preparation and characterization adjacent to the beamlines. A proactive maintenance plan ensures the high level of performance and minimizes downtown. A major upgrade of beamline optics, endstation, and computing resources will prepare GM/CA for the planned 2023 arrival of a new storage ring source known as APS-U. The new optics will enable a focused, 1-micron beam, and the new source will increase brightness by 100-fold compared to the current APS. Outreach activities, user workshops, and new user training materials will prepare the user community for the opportunities and challenges of APS-U.

GM/CA@APS 资源的主要目标是为用户提供对尖端技术的访问 大分子晶体学光束线确定前沿问题的 3D 原子结构 该资源成立于 1999 年，并作为国家用户运行。 自 2004 年以来一直致力于为大分子晶体学提供全套尖端技术 阿贡国家实验室高级光子源 (APS) 23 区由 GM/CA 领导。 Robert Fischetti（阿贡国家实验室，联系人 PI）和 Janet Smith（密歇根大学，PI） 拥有有效的管理结构来操作、维护和升级光束线，并运行 高影响力的用户程序。 GM/CA 大分子晶体学核心提供了现代、 强大的环境，配备先进的样本筛选、询问和数据收集工具 通过两个插入装置 X 射线束线 23ID-B 的操作实现固定能量、SAD 和 MAD 模式 和 23ID-D，来自倾斜波荡器源，光束线可在较宽范围内独立调谐。 能量范围，旨在提供位置和强度极其稳定的明亮 X 射线束。 卓越的稳定性使 GM/CA 成为异常散射的理想工具，用户可以快速 （几分钟）在 5.0-20 keV 范围内随意改变能量 (λ = 2.48 Å - 0.62 Å) 稳定光束。 实现了强大的微晶体学能力。 四微型光束准直器，用户可选光束尺寸为 5、10 和 20 微米直径 除了全光束之外，用户还可以随时快速（几秒钟内）更改任何样品的光束尺寸。 光束线配备了快速、低噪声的像素阵列探测器， 23ID-B 上的 Eiger 16M 和 23ID-D 上的 Pilatus3 6M 均由机器人安装。 来自 288 个样本的杜瓦瓶，可以在光束中以微米精度进行可视化和操作。 用户通过直观且生物学家友好的 JBluIce 图形用户控制光束线和样品 JBluIce 包括基于 GUI 的光束线控制、样品居中、样品功能。 筛选、数据收集、X射线荧光能量扫描、样品光栅扫描和数据分析。 强大的光栅功能用于扫描微晶体场或找到最佳衍射区域 来自光栅扫描的信息可以与各种数据收集例程共享。 数据收集选项提供晶胞和分辨率估计。 制定数据收集策略，自动耦合到三个数据处理管道， 2016-2018 年期间，总共为用户提供了近乎实时的反馈 857 个。 来自 157 个独特群体的个人使用 GM/CA 光束线，每年询问 35,000 个样本， 在 PDB 中存放了 225 个结构，并发表了 130 篇论文（12% 发表在《自然》、《科学》和《细胞》上）。 实施串行晶体学方法并将其集成到 JBluIce 自动化数据中 用于粘性注射器和固定目标样品输送的收集和处理管道 计算集群和存储阵列网络提供高速数据存储、检索、 用户接受专家人员的培训，并可以收集数据。 工作人员还为各种社区外展活动做出贡献，包括现场或远程。 年度 CCP4/APS 晶体学学校 生物化学实验室提供样品的基本功能。 邻近光束线的准备和表征 主动维护计划可确保 高水平的性能并最​​大限度地减少市中心的光束线光学器件、终端站和系统的重大升级。 计算资源将为 GM/CA 计划于 2023 年到来的已知新存储环源做好准备 作为 APS-U，新的光学器件将实现聚焦的 1 微米光束，并且新的光源将增加 与当前的 APS 外展活动、用户研讨会和新用户相比，亮度提高了 100 倍。 培训材料将使用户社区为 APS-U 的机遇和挑战做好准备。