Administrative Supplement for Structural Dynamics in Biology Resource Year 2

生物资源第二年结构动力学行政补充

• 批准号: 10833964
• 负责人: Sebastien Boutet
• 金额: $ 18.9万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10833964
• 关键词: 3D Print; Administrative Supplement; Automobile Driving; Behavior; Biological; Biological Process; Biology; Cardiovascular Diseases; Ceramics; Characteristics; Communities; Complement; Cryopreservation; Crystallography; Data Collection; Decision Making; Development; Devices; Drug Targeting; Electron Microscopy Facility; Ensure; Exposure to; G-Protein-Coupled Receptors; Geometry; Goals; Integral Membrane Protein; Laboratories; Light; Liquid substance; Malignant Neoplasms; Maps; Measurement; Measures; Mental disorders; Metalloproteins; Metals; Methods; Microfluidics; Oxidation-Reduction; Physiologic pulse; Plant Resins; Positioning Attribute; Predisposition; Process; Property; Publishing; Radiation; Radiation induced damage; Resolution; Resources; Roentgen Rays; Sampling; Signal Transduction; Solvents; Source; Speed; Structure; Synchrotrons; System; Technology; Time; Training; Translating; Viscosity; Work; base; biomaterial compatibility; density; detector; drug development; experimental study; fabrication; metalloenzyme; novel; open source; parent grant; programs; prototype; radiation mitigation; structural biology; therapeutic development

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.

项目摘要 - 行政补充请求 - 1P41GM139687-02 - Sebastien boutet（pi）。 这种行政补充要求是由整体目标和结构的目标驱动的 SLAC国家加速器实验室生物学BTRR的动力学。 BTRR针对 增强和开发SLAC LINAC相干光源的独特功能 （LCL）用于生物医学应用。请求的补充剂将支持广泛的用户群 进一步进一步的DBP和TR＆D的目标。 通过克服同步基因辐射损伤的局限性，LCLS尤其是 在大型大分子机器的研究中产生影响 内容，使其变得精致，难以冷冻的良好，并且对辐射非常敏感。为了 例如，主要突破是SFX的应用来检查固有的晶体 膜蛋白，例如GPCR，在LCP生长。 GPCR是最大的目标群 药物开发用于治疗多种疾病（例如癌症，心血管疾病， 和精神疾病）。 LCLS在对金属酶的研究中也具有影响，这对于 几乎所有的生物过程，因此代表了丰富的治疗目标空间 发展。金属蛋白的高分辨率结构研究在 同步加速器是因为金属中心，尤其是氧化还原活性的中心，非常非常 易受X射线诱导的光导量的影响。此外，产生的超快（〜40 fs）X射线脉冲 通过LCLS开放新的可能性，直接观察涉及的动态过程 大分子功能。此外，许多P41衍生的发展使得能够快速 使用多个小晶体收集数据，既适用于LCLS和同步加速器 减轻辐射损伤。通过扩展LCLS和SSRL同步器的功能， BTRR为结构表征打开更多的大分子机器，包括时间 在广泛的生物医学相关时间尺度上解决的研究。与和 增强SSRL和LCLS的现有程序，BTRR将提供支持，专业知识 并培训了广泛的生物医学界。 这种行政补充剂将增强和扩展提供的BTRR功能 向普通用户和p41驱动生物医学项目。获得高速X射线 切菜器将满足BTRR的迫切需要，以有效利用SSRL BL12-1与小晶体 由液体/晶体喷油器和时间分辨测量。除了实现 微秒时间分辨率，通过将连续的X射线束分解为短脉冲， 切碎机确保注射器输送的晶体不会在X射线上破坏损坏 完全翻译成X射线梁位置，使暴露于未损坏 BL12-1时单色或粉红色梁。此外，该补充要求具有生物相容性 树脂和陶瓷3D打印机具有2微米分辨率，将允许快速原型制作和 优化样品喷油器，可以提供可靠性和易度性，以扩大用户 社区。最后，该提案还要求系统来测量样品的电导率 以自动化的方式，将允许进一​​步表征BTRR的目标 条件并将其映射到理想的样品输送方法上，以用于给定 样本，为用户提供有关如何最好地执行实验的结构性决策。 通过增加电导率， 电子样品传递的一个重要参数，以发布查找表。