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-专业高分子结晶学 该技术运营核心将提供九项先进技术，这些技术是 致力于解决特别具有挑战性的问题的结构生物学用户社区的成员。这些是：1) 单晶不够用时的多晶策略，2)制备时的自然元素分相 衍生品是不切实际的，3)当晶体太脆弱而无法处理时，来自托盘的原位衍射，4) 非低温下的衍射，用于功能研究或低温保护时的衍射 更糟糕的是，另一种可视化技术用于在从偏振范围内的环路中寻找晶体 显微镜到在线X射线层析成像(CBOXAR)和对小X射线束的栅格搜索 对样品表面的衍射质量进行详尽的探测，6)基于一阶的数据质量预测。 推导最佳数据采集参数的原理和大尺度衍射模拟技术 基于关于给定样本的所有可用信息，7)可用波束的综合阵列 性能，包括我们即将完成的微聚焦双子座光束线自动光学重新定心 技术(AUORA)，支持将实验自主迁移到任何ALS光束线，以进行优化 光束线利用9)明确的“目标文件格式”规范，以便在必要时，可以进行实验 迁移到肌萎缩侧索硬化症之外，例如X射线自由电子激光。 所有这些技术都将通过支持肌萎缩侧索硬化症的网站捆绑在一起，该网站不仅追踪样本 和数据处理结果，但它们之间的相互兼容关系。这将是至关重要的 管理数据集的组合爆炸，必须探索这些数据集才能以最好的方式组合在一起 给定项目的完整数据集。这一平均将是本机元素相位的关键，其中信号 从任何单个样本中提取的数据都不足以用于阶段化。该网站还将作为一个知识库， 能够根据用户当前拥有的所有数据和预测向用户提供建议 由我们独一无二的精确衍射模拟技术提供。例如，建议您 如果SAXS(TOC2)表明他们的分子本质上是有序的，但衍射是有序的，他们就会尝试原位衍射 停留在6？分辨率。如果原位衍射性也差，那么建议寻找一种新的 使用X射线足迹(ALS使能外部)或低温EM(ALS外部)形成晶体。 我们预计该资源将吸引广泛的区域和国家地理分布的用户。通过 解决衍射差的问题，需要在多个温度下进行功能研究，需要 对于原生元素的阶段化，通过分散晶体中心问题，我们将利用多样性 将ALS波束线转换成连贯且易于访问的资源。