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

基本信息

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
项目类别：
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

项目摘要

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 布泰（PI）。该行政补充请求是由结构性组织的总体目标和目标驱动的 SLAC 国家加速器实验室的生物学动力学 BTRR BTRR 的目标是。增强和开发 SLAC 直线加速器相干光源的独特功能（LCLS）用于生物医学应用。所请求的补充将支持广泛的用户群。并进一步实现 DBP 和 TR&D 的目标。通过克服同步加速器辐射损伤的局限性，LCLS 特别适用于对研究用高溶剂形成小晶体的大分子机器具有影响力含量，使得它们脆弱、难以冷冻保存并且对辐射极其敏感。例如，一项重大突破是应用 SFX 来检查本征晶体在 GPCR 中生长的膜蛋白（例如 GPCR）是最大的靶标群体。药物开发，用于治疗多种疾病（例如癌症、心血管疾病、 LCLS 在金属酶的研究中也有影响，这对金属酶的研究至关重要。几乎所有的生物过程，因此代表了丰富的治疗目标空间金属蛋白的高分辨率结构研究尤其具有挑战性。同步加速器，因为金属中心，特别是那些具有氧化还原活性的金属中心，非常此外，容易受到 X 射线诱导的光还原作用，产生超快 (~40 fs) X 射线脉冲。 LCLS 为直接观察涉及的动态过程开辟了新的可能性此外，许多 P41 衍生的发展使得快速发展成为可能。使用多个小晶体收集数据，适用于 LCLS 和同步加速器通过扩展 LCLS 和 SSRL 同步加速器的能力来减轻辐射损伤 BTRR 为更多的大分子机器提供了结构表征，包括时间- 解决了广泛的生物医学相关时间尺度的研究。 BTRR 将提供支持和专业知识，以加强 SSRL 和 LCLS 的现有计划以及对广大生物医学界的培训。该行政补充将增强和扩展所提供的 BTRR 功能普通用户和 P41 驱动生物医学项目高速 X 射线的采集。斩波器将满足 BTRR 高效使用 SSRL BL12-1 和小晶体的关键需求除了实现之外，还可以通过液体/晶体注射器提供并进行时间分辨测量。微秒时间分辨率，通过将连续的 X 射线束分解成短脉冲，斩波器可确保注射器输送的晶体在被 X 射线损坏之前不会被破坏完全转换为 X 射线束位置，从而能够在无人值守的情况下进行曝光 BL12-1 为单色或粉红色光束此外，该补充剂需要生物相容性。具有 2 微米分辨率的树脂和陶瓷 3D 打印机，可实现快速原型制作和进样器的优化将提供可靠性和易用性，从而扩大用户范围最后，该提案还要求建立一个测量样品电导率的系统。以自动化方式进一步实现 BTRR 在表征样本方面的目标条件并将其映射到用于给定的理想样品输送方法示例，为用户提供有关如何最好地执行实验的结构化决策。这将通过增加电导率来补充物理化学表征方面正在进行的努力，动电样品输送的一个重要参数，用于查找要发布的表。