RAPID Enhanced SARS-CoV-2 High-Throughput Crystallization for Structural Studies

该奖项授予Hauptman-Woodward研究所，以扩大其结构生物学服务，为研究SARS-CoV-2相关样本的研究人员提供服务，并加速SARS-CoV-2病毒蛋白的知识生产。结构生物学方法有助于理解生物分子如何工作的基础上的基本原理，使这些分子的可视化在一个高层次的细节。自从SARS-CoV-2在2019年底成为一种致病性威胁以来，结构生物学方法在提供有关病毒如何进入人类细胞以及病毒蛋白如何与人类蛋白相互作用的细节方面已经变得至关重要。实现这些结构图的一个困难步骤是定位允许研究蛋白质样品的实验条件，而该项目旨在解决这一瓶颈。结晶中心为世界各地研究蛋白质结构的结构生物学研究人员提供最先进的机器人设备和专业成像。该项目将提供对这些资源的即时访问，并为直接处理SARS-CoV-2相关样本的研究人员开发更好的服务。该项目还将开发新的实验管道，以加快应对当前和未来大流行病的时间。影响人类社会的病毒数量持续增加，最近爆发的SARS、MERS、埃博拉和寨卡病毒等就是证明。病毒有可能破坏国家的健康和繁荣，因此，像这项研究这样的努力有可能有助于解决这些全球性问题。自从新病原体SARS-CoV-2出现以来，已有超过100种病毒蛋白结构被存入蛋白质数据库，其中包括92种最近沉积的SARS-CoV-2主要蛋白酶与不同配体的复合物（截至2020年4月15日）。 到目前为止，近95%的沉积结构来自大分子X射线晶体学（MX）实验。MX实验的主要瓶颈之一是确定蛋白质结晶的条件，这阻碍了当前流行病所需的快速反应。结晶中心是一个高通量结晶设施，促进生物分子靶点的快速，有效结晶，是世界上唯一可供外部用户使用的资源，使高通量结晶筛选在一个实验板中具有1，536个条件，并提供最先进的成像方法，能够非常快速地检测蛋白质晶体。这些资源正在提供给研究SARS-CoV-2样本的研究人员。此外，结晶管道正在扩大，1）增加实验管道中最先进的成像，2）在设施中进行优化和放大实验，以及3）加快晶体样品的收获和运输。该项目将对科学界快速解决SARS-CoV-2蛋白质结构的能力产生深远影响，促进对冠状病毒生物学基础科学的更好理解。该RAPID奖由生物基础设施部（DBI）使用冠状病毒援助，救济，该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

An award is made to Hauptman-Woodward Institute to extend its structural biology services for researchers working with SARS-CoV-2 related samples and to accelerate knowledge production about SARS-CoV-2 virus proteins. Structural biology methods help in understanding fundamental principles at the base of how biomolecules work by enabling visualization of those molecules at a high level of detail. Since SARS-CoV-2 emerged as a pathogenic threat in late 2019, structural biology methods have already been critical in providing details about how the virus enters human cells and how viral proteins interact with human proteins. One difficult step in achieving these structural maps is locating experimental conditions that allow the protein samples to be studied, and this project is geared toward addressing that bottleneck. The Crystallization Center provides state-of-the-art robotic equipment and specialized imaging to structural biology researchers worldwide who are studying protein structures. This project will provide immediate access to these resources and enable enhanced services to be developed for researchers working directly with SARS-CoV-2 related samples. This project will also develop new experimental pipelines to accelerate response time in the face of the current and future pandemics. The number of viruses that impact human society continues to rise, as evidenced by recent outbreaks of SARS, MERS, Ebola and Zika virus, among others. Viruses have the potential to damage national health and prosperity, and efforts such as this research therefore have potential to contribute to solutions to these global problems. Since the appearance of the novel pathogen SARS-CoV-2, over 100 structures of viral proteins have been deposited to the Protein Data Bank, including 92 recent depositions of the SARS-CoV-2 main protease in complex with different ligands (as of April 15, 2020). Close to 95% of the structures deposited thus far are derived from macromolecular X-ray crystallography (MX) experiments. One of the major bottlenecks in MX experiments is determining the conditions in which a protein will crystallize, and this hinders the rapid response needed in the current pandemic. The Crystallization Center is a high-throughput crystallization facility that facilitates rapid, efficient crystallization of biomolecular targets and is the only resource in the world available to external users that makes high-throughput crystallization screening accessible with 1,536 conditions in one experimental plate and provides state-of-the-art imaging methods capable of detecting protein crystals very rapidly. These resources are being made available to researchers working with SARS-CoV-2 samples. Additionally, crystallization pipelines are being expanded with 1) increased state-of-the-art imaging in the experimental pipeline, 2) optimization and scale-up experiments at the facility, and 3) expedited crystal sample harvesting and shipping. This project will have a profound impact on the scientific community’s ability to quickly solve SARS-CoV-2 protein structures, facilitating a better understanding of the basic science underlying coronavirus biology.This RAPID award is made by the Division of Biological Infrastructure (DBI) using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.