Administrative Supplement for Structural Dynamics in Biology Resource Year 2

项目摘要-行政补充申请-1P41GM139687-02-Sebastien 布特(PI)。 这一行政补充请求是由结构性的总体目标和目的驱动的 SLAC国家加速器实验室生物动力学BTRR。BTRR的目标是 增强和发展SLAC直线加速器相干光源的独特能力 (LCLS)用于生物医学应用。所要求的补充内容将支持广泛的用户基础 并推进DBP和R&D的目标。 通过克服同步加速器辐射损害的限制，LCLS特别 在形成高溶剂小晶体的大分子机器的研究中具有影响力 这使它们变得脆弱，很难冷冻保存，而且对辐射非常敏感。为 例如，一项重大突破是应用SFX来检查本征晶体 LCP中生长的膜蛋白，如GPCRs。GPCR是全球范围内 药物开发，用于治疗各种疾病(例如癌症、心血管疾病、 和精神疾病)。LCLS在金属酶的研究中也很有影响力，金属酶对 几乎所有的生物过程都代表着治疗学的丰富靶点空间 发展。金属蛋白的高分辨率结构研究在以下方面尤其具有挑战性 同步加速器是因为金属中心，特别是那些氧化还原活跃的中心，非常 易受x射线诱导的光还原。此外，产生的超快(~40飞秒)X射线脉冲 通过LCLS为直接观察涉及的动态过程打开了新的可能性 大分子功能。此外，许多从P41派生的发展使快速 使用多个小晶体收集数据，适用于LCLS和同步加速器 以减轻辐射损害。通过扩展LCLS和SSRL同步加速器的能力， BTRR开放了更多的大分子机器来进行结构表征，包括时间- 解决了广泛的生物医学相关时间范围内的研究。与和集成 加强SSRL和LCLS的现有计划，BTRR将提供支持、专业知识 以及对广大生物医学界的培训。 这一行政补充将增强和扩展提供的BTRR能力 面向普通用户和推动生物医学项目的P41。高速X光机的获取 斩波将满足BTRR有效使用具有小晶体的SSRL BL12-1的关键需求 由液晶/晶体注射器提供，用于时间分辨测量。除了实现 微秒时间分辨率，通过将连续的X射线束分解成短脉冲， 切割器确保注射器输送的晶体在被X射线破坏之前不会被破坏 被完全转换到x射线束位置，从而能够暴露在未衰减的 BL12-1的单色或粉色光束。此外，这种补充剂要求具有生物相容性 树脂和陶瓷3D打印机，分辨率为2微米，可实现快速原型制作和 样品注射器的优化将提供可靠性和易用性，从而扩大用户范围 社区。最后，该提案还要求建立一个系统来测量样品的电导率 以自动化的方式实现BTRR在表征样品方面的目标 并将这些条件映射到理想的样品传递方法上，以用于给定的 示例，为用户提供关于如何最好地执行他们的实验的结构化决策。 这将通过增加导电性来补充正在进行的物理化学表征方面的努力， 电动样品传递的一个重要参数，以查找表即将发布。

Project Summary – Administrative Supplement Request – 1P41GM139687-02 – Sebastien Boutet (PI). This administrative supplement request is driven by overall goals and aims of the Structural Dynamics in Biology BTRR at SLAC National Accelerator Laboratory. The BTRR is aimed at enhancing and developing the unique capabilities of the SLAC Linac Coherent Light Source (LCLS) for biomedical applications. The requested supplement will support a broad user base and further the goals of the DBPs and the TR&Ds. By overcoming the limitations of radiation damage at the synchrotron, LCLS has been particularly impactful in the study of large macromolecular machines that form small crystals with high solvent content, making them delicate, difficult to cryo-preserve and extremely radiation sensitive. For example, a major breakthrough was the application of SFX to examine crystals of intrinsic membrane proteins, such as GPCRs, grown in LCP. GPCRs are the largest group of targets for drug development, used to treat a wide variety of illnesses (e.g. cancer, cardiovascular disease, and mental illness). LCLS is also impactful in the study of metalloenzymes, which are critical to nearly all biological processes and as such represent a rich target space for therapeutics development. High-resolution structural studies of metalloproteins are particularly challenging at the synchrotron because the metal centers, especially those that are redox active, are very susceptible to x-ray induced photoreduction. Further, the ultrafast (~40 fs) x-ray pulses produced by the LCLS open new possibilities in directly observing dynamic processes involved in macromolecular function. Moreover, many of the P41-derived developments that enable the rapid collection of data using multiple small crystals, are applicable both at LCLS and at the synchrotron to mitigate radiation damage. By expanding the capabilities at LCLS and at the SSRL synchrotron, the BTRR opens more macromolecular machines to structural characterization, including time- resolved studies over a wide range of biomedically relevant time scales. Integrating with, and enhancing the existing programs at SSRL and LCLS, the BTRR will provide support, expertise and training to the broad biomedical community. This administrative supplement will enhance and expand the BTRR capabilities that are provided to general users and to the P41 driving biomedical projects. The acquisition of a high speed x-ray chopper will fill a critical need of the BTRR for efficient use of SSRL BL12-1 with small crystals delivered by liquid/crystal injectors and for time-resolved measurements. In addition to achieving microsecond time resolution, by breaking up the continuous x-ray beam into short pulses, the chopper ensures crystals delivered by injectors are not destroyed by x-ray damage before they are fully translated into the x-ray beam position, enabling exposure to unattenuated monochromatic or pink-beam at BL12-1. In addition, this supplement requests a biocompatible resin and ceramic 3D printer with 2-micron resolution that will allow for rapid prototyping and optimization of sample injectors that will provide reliability and ease that will broaden the user community. Finally, the proposal also requests a system to measure the conductivity of samples in an automated manner which will allow to further the goals of the BTRR in characterizing sample conditions and mapping those onto the ideal sample delivery method to be used for a given sample, providing users with structured decision-making on how to best perform their experiment. This will complement ongoing efforts in physicochemical characterization by adding conductivity, an important parameter for electrokinetic sample delivery, to lookup tables to be published.