RAPID Collaborative Proposal: High-resolution structure determination of Coronavirus ligands by high-throughput WAXS

RAPID 合作提案：通过高通量 WAXS 测定冠状病毒配体的高分辨率结构

• 批准号: 2030052
• 负责人: Diana Monteiro
• 金额: $ 20万
• 依托单位: Hauptman-Woodward Medical Research Institute Inc
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2030052&HistoricalAwards=false
• 关键词: RAPID; Collaborative; Proposal; resolution; structure

This RAPID award is made by the Division of Biological Infrastructure (DBI) using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act. The award to Hauptman-Woodward Medical Research Institute supports research with a novel strategy that yields structural information on protein interactions in solution and with high-throughput by using x-ray scattering techniques. The researchers will use their rapid analyses to understand how proteins critical to SARS-COV-2 viral replication function within cells of infected individuals, and how these proteins interact with potential therapeutic drug compounds. High-throughput, rapid hit detection and fast structure solutions are essential for a timely and aggressive response to this pandemic. Results from these studies will contribute vital information to our understanding of SARS-COV-2 biology and may help to inform on the selection and development of therapeutic treatments. Students from under-represented groups will also participate in this activity through the BioXFEL Scholars program to help with data analysis. Data from these studies will be rapidly disseminated through publicly available repositories so that other researchers can leverage the experimental outcomes in their own studies. The study findings will also be published in peer-reviewed journals and shared at scientific meetings.This project will use small and wide-scattering X-ray techniques (SWAXS) to deliver a large database of structural information on protein-ligand interactions for the NSP5 protease and then NSP13 helicase (and potentially NSP15), including binding-induced structural changes. The successful application of high-throughput SWAXS screening approach to SARS-CoV-2 proteins will provide the community with valuable structural information on molecular interactions modulating viral function. These studies will focus initially on the ligand binding with the main viral protease, NSP5, responsible for the release of various proteins encoded in the large polyprotein sequence. Analyses will also be performed to investigate NSP13, a protein known to interact with many human proteins involved in the innate immune response. Finding surface binding hotspots will provide information on potential binding interfaces as well as initial small molecule scaffolds that future efforts can use to target these interfaces.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.

这一快速奖励是由生物基础设施部(DBI)利用冠状病毒援助、救济和经济安全(CARE)法案的资金做出的。该奖项授予Hauptman-Woodward医学研究所，支持采用一种新策略的研究，该策略通过使用X射线散射技术产生关于溶液中蛋白质相互作用的结构信息和高通量。研究人员将利用他们的快速分析来了解对SARS-COV-2病毒复制至关重要的蛋白质在感染者细胞内的功能，以及这些蛋白质如何与潜在的治疗药物化合物相互作用。高吞吐量、快速命中检测和快速结构解决方案对于及时和积极地应对这一流行病至关重要。这些研究的结果将有助于我们了解SARS-COV-2生物学，并可能有助于为治疗方法的选择和开发提供信息。来自代表性不足群体的学生也将通过BioXFEL学者计划参加这项活动，以帮助进行数据分析。这些研究的数据将通过公开可用的储存库迅速传播，以便其他研究人员可以在他们自己的研究中利用实验结果。这项研究成果还将发表在同行评议的期刊上，并在科学会议上分享。该项目将使用小而宽散射X射线技术(SWAXS)来提供一个大型数据库，其中包括NSP5蛋白酶和NSP13解旋酶(可能还有NSP15)的蛋白质-配体相互作用的结构信息，包括结合诱导的结构变化。成功地将高通量SWAXS筛选方法应用于SARS-CoV-2蛋白，将为社区提供关于调控病毒功能的分子相互作用的有价值的结构信息。这些研究最初将集中在与主要病毒蛋白酶NSP5结合的配体上，NSP5负责释放在大的多蛋白序列中编码的各种蛋白质。还将进行分析以研究NSP13，一种已知与许多参与先天免疫反应的人类蛋白相互作用的蛋白质。发现表面结合热点将提供关于潜在结合界面以及未来工作可以用来针对这些界面的初始小分子支架的信息。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Plug-and-play polymer microfluidic chips for hydrated, room temperature, fixed-target serial crystallography.

• DOI: 10.1039/d1lc00810b
• 发表时间: 2021-12-07
• 期刊: Lab on a chip
• 影响因子: 6.1
• 作者: Gilbile D;Shelby ML;Lyubimov AY;Wierman JL;Monteiro DCF;Cohen AE;Russi S;Coleman MA;Frank M;Kuhl TL
• 通讯作者: Kuhl TL