Specialized Macromolecular Crystallography

专业高分子晶体学

• 批准号: 10201650
• 负责人: James M Holton
• 金额: $ 36.42万
• 依托单位: UNIVERSITY OF CALIF-LAWRENC BERKELEY LAB
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-09-20
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201650
• 关键词: Address; Award; Back; Binding; Biological Assay; Blood capillaries; Characteristics; Communities; Cryoelectron Microscopy; Crystallization; Crystallography; Data; Data Collection; Data Set; Defect; Diagnostic Procedure; Disease; Dose; Dose-Rate; Effectiveness; Elements; Explosion; Face; Feedback; Geographic Distribution; Harvest; In Situ; Infrastructure; Ligands; Logistics; Maintenance; Maps; Methods; Molecular Conformation; Mutation; Optics; Phase; Philosophy; Polarization Microscopy; Polymers; Property; Proteins; Radiation induced damage; Recommendation; Resolution; Resources; Roentgen Rays; Route; Sampling; Shapes; Signal Transduction; Sleep; Source; System; Technology; Temperature; Thick; Time; Visualization; Work; X-Ray Tomography; base; beamline; combinatorial; computerized data processing; cost; data quality; design; exhaustion; experimental study; file format; improved; knowledge base; member; migration; operation; optical imaging; screening; simulation; structural biology; web site; x-ray free-electron laser

Project Summary/Abstract - Core 3 – Specialized Macromolecular Crystallography This Technology Operations Core will serve up nine advanced technologies that are seriously needed by members of the structural biology user community working on particularly challenging problems. These are: 1) multi-crystal strategy for when one crystal is not enough, 2) native element phasing for when preparing derivatives is impractical, 3) in-situ diffraction from trays for when the crystals are too fragile to handle, 4) diffraction at non-cryogenic temperatures, for functional studies or when cryo-protection makes diffraction worse, 5) alternative visualization technologies for finding crystals in loops ranging from polarization microscopy to online X-ray tomography (CBOXAR) and raster grid searches of a small x-ray beam over the face of the sample to probe for diffraction quality exhaustively, 6) data quality prediction based on first- principles and at-scale diffraction simulation technology to deduce the best possible data collection parameters based on all available information about a given sample, 7) a comprehensive array of available beam properties, including our soon-to-be-completed micro-focus GEMINI beamline 8) automatic optical re-centering technology (AUORA) to enable autonomous migration of experiments to any ALS beamline, for optimizing beamline utilization 9) a clear “targeting file format” specification so that, if necessary, experiments can be migrated outside the ALS, such as to X-ray Free Electron Lasers. All these technologies will be tied together by the ALS-ENABLE website, which will track not just the samples and data processing results, but the inter-compatibility relationships between them. This will be essential for managing the combinatorial explosion of data sets that must be explored to stitch together the best possible complete data set for a given project. This averaging will be key to native-element phasing, where the signal from any single sample is seldom good enough for phasing. The website will also serve as a knowledge base, capable of making recommendations to Users based on all the data they currently have, and the predictions provided by our uniquely accurate diffraction simulation technology. For example, it will be recommended that they try in-situ diffraction if SAXS (TOC 2) shows that their molecule is intrinsically ordered but diffraction is stuck at 6 Å resolution. If in-situ diffraction is also poor, then the recommendation will be to search for a new crystal form using X-ray Footprinting (outside ALS-ENABLE) or cryo-EM (outside ALS). We expect this Resource will appeal to a wide regional and national geographic distribution of users. By addressing the problem of poor diffraction, the need for functional studies at multiple temperatures, the need for native element phasing, and by de-centralizing the crystal centering problem we will leverage the diversity of the ALS beamlines into a coherent and easily accessible Resource.

项目摘要/摘要 - 核心3 - 专门的大分子晶体学 该技术运营核心将为九种高级技术服务，这些技术非常需要 结构生物学用户社区的成员致力于特别具有挑战性的问题。这些是：1） 多结晶策略对于一个晶体不够，2）本机元素逐步逐步进行准备 衍生物是不切实际的，3）托盘的原位衍射，因为晶体太脆弱而无法处理，4） 在非晶体温度下，用于功能研究或冷冻保护使衍射时的衍射 更糟糕的是，5）在偏振范围内寻找晶体的替代可视化技术 在线X射线断层扫描（Cboxar）的显微镜和对小X射线梁的栅格网格搜索 样品面对以详尽的探测衍射质量，6）基于先前的数据质量预测 原理和尺度衍射模拟技术，以推导最佳的数据收集参数 基于有关给定样品的所有可用信息，7）一系列可用的光束 属性，包括我们即将完成的微焦点双子座光束线8）自动重新介入 技术（AUORA）可以使实验自主迁移到任何ALS梁线，以优化 光束线利用率9）明确的“针对文件格式”规范，以便在必要时可以实验 迁移在ALS之外，例如X射线无电子激光器。 所有这些技术都将由ALS-Enable网站捆绑在一起，该网站不仅会跟踪样品 和数据处理结果，但它们之间的兼容关系。这对于 管理必须探索必须探索的数据集的组合爆炸，以将最佳缝合在一起 给定项目的完整数据集。这个平均值将是本地元素相位的关键 从任何单个样品中，很少有足够的好处来进行计量。该网站还将作为知识库， 能够根据他们当前拥有的所有数据向用户提出建议，预测 由我们独特的准确衍射模拟技术提供。例如，建议 如果SAXS（TOC 2）表明它们的分子是本质上的，但衍射为 粘在6Å分辨率。如果原位衍射也很差，则建议是搜索新的 使用X射线足迹（外部ALS-Enable）或Cryo-Em（外部ALS）的晶体形式。 我们希望这种资源将出现在用户的区域和国家地理分布中。经过 解决衍射差不良问题，需要在多个温度下进行功能研究的需求 对于本地元素阶段，通过偏心晶体居中问题，我们将利用多样性 ALS光束线成相干且易于访问的资源。