X-ray Crystallographic Fragment Screening Core

X射线晶体碎片筛选核心

• 批准号: 10363021
• 负责人: EDWARD ARNOLD
• 金额: $ 39.4万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10363021
• 关键词: ray; Crystallographic; Fragment; Screening; Core

Abstract X-ray crystallographic fragment screening (XCFS) is a powerful tool for identification of new pharmacophores for probes and drug design as well as well as for biochemical and structural dynamic characterization of the protein of interest. An XCFS Core has been established to make this powerful tool accessible to multiple HIVE Projects studying HIV-1 and related proteins/complexes in the HIVE. The XCFS Core (Arnold group) has completed three XCFS campaigns against the viral targets: HIV-1 reverse transcriptase, influenza A endonuclease, and HIV-1 integrase catalytic core domain. In each case the successful screening campaign has discovered novel hits and in the cases of HIV-1 reverse transcriptase and influenza endonuclease has found novel conformations/pockets for understanding protein dynamics and biochemistry. The XCFS Core will collaborate with the other HIVE Projects and Cores to identify crystal forms that are appropriate for XCFS and optimize them as needed. Initial projects include: HIV-1 capsid (Project 3, Sarafianos), HIV-1 integrase (Projects 1 and 4, Kvaratskhelia and Engelman), and HIV-1 reverse transcriptase with nucleic acid substrate (Project 3, Arnold). Small pilot screening will verify the usefulness of the crystal form for XCFS before proceeding to large scale screening using very high-throughput synchrotron X-ray crystallography beamlines (Specific Aim 1). A specialized beamline, XChem, at the Diamond Light Source will be used to efficiently and rapidly screen 1000-2000 small molecule fragments. The XChem facility has developed several technologies for fragment soaking, crystal handling, data collection and processing to allow for very rapid XCFS campaigns. In addition to XChem, there are new high-speed beamlines at Brookhaven National Laboratories and elsewhere capable of data-collection speeds unobtainable two years ago. The new beamline technologies allow for fragments from a large library to be screened individually, whereas previously mixtures of fragments needed to be used. The ability to screen individual compounds allows for design of novel compound libraries to take advantage of this new technology (Specific Aim 2). New fragment libraries with click-reactive and SuFEx-containing moieties will be designed for XCFS (Project 6, Olson and Sharpless). Data from XCFS will be processed and refined to ensure accurate hit identification and hits will be verified through screening of derivatives and appropriate orthogonal assays. Communication and data management between the Projects and Cores will ensure clear information transfer and follow up chemistry/studies (Specific Aim 3).

抽象的 X射线晶体学片段筛选（XCFS）是识别新药算术的强大工具 用于探针和药物设计以及生化和结构动态表征 感兴趣的蛋白质。已经建立了XCFS核心，以使该功能强大的工具可访问多个Hive 研究Hive中研究HIV-1及相关蛋白质/复合物的项目。 XCFS Core（Arnold Group）具有 完成了针对病毒靶标的三个XCF运动：HIV-1逆转录酶，流感A 核酸内切酶和HIV-1整合酶催化核心结构域。在每种情况下，成功的筛选活动 在HIV-1逆转录酶和流感核酸内切酶的情况下，发现了新颖的热门单曲 找到了用于了解蛋白质动力学和生物化学的新型构象/口袋。 XCFS Core将与其他Hive项目和内核合作，以识别晶体形式 适用于XCF，并根据需要进行优化。初始项目包括：HIV-1 CAPSID（项目3， Sarafianos），HIV-1积分酶（项目1和4，Kvaratskhelia和Engelman）和HIV-1反向 带有核酸底物的转录酶（项目3，阿诺德）。小型飞行员筛选将验证 XCF的晶体形式在使用非常高通量同步加速器进行大规模筛选之前 X射线晶体学束线（特定目标1）。钻石灯处的专门光束线Xchem 来源将用于有效，快速筛选1000-2000个小分子片段。 Xchem设施 已经开发了几种用于碎片浸泡，水晶处理，数据收集和处理的技术 允许非常快速的XCFS广告系列。除Xchem外，还有新的高速梁线 Brookhaven国家实验室和其他能够数据收集速度无法实现的两年 前。新的光束线技术允许单独筛选大型库的片段， 而先前需要使用碎片的混合物。筛选单个化合物的能力 允许设计新颖的复合库可以利用这项新技术（特定的目标2）。新的 具有点击反应和含Sufex的部分的碎片库将是为XCF设计的（项目6，项目6， Olson和Sharpless）。 XCF的数据将进行处理和精制，以确保准确的命中识别并将验证命中率 通过筛选衍生物和适当的正交测定。通信和数据管理 在项目和核心之间将确保清晰的信息转移并跟进化学/研究（具体 目标3）。