RAPID: IIBR: Coronavirus Viroporin XFEL Structural Studies

目前迫切需要确定严重急性呼吸综合征冠状病毒-2 （SARS-CoV-2）蛋白的结构。冠状病毒是包膜病毒，由四种主要结构蛋白组装而成：膜(M)、刺突(S)、包膜(E)和核衣壳(N)。该奖项支持的研究人员将重点研究冠状病毒E蛋白，以了解这些蛋白如何通过与细胞膜的相互作用，在病毒基因组被包裹时促进出芽过程。这些研究将有助于识别和设计用于结构-功能研究的分子，并可能提供对治疗方法设计至关重要的结构信息。博士后和研究生学员，包括来自代表性不足群体的成员，将全面参与研究工作。这些研究的结果将发表在同行评议的期刊上，在科学会议上提出，并通过共享数据库提供。该奖项支持的研究目标是解决高致病性人类冠状病毒的冠状病毒E蛋白的原子结构，这些冠状病毒包括中东呼吸综合征（MERS）、严重急性呼吸综合征（SARS）和最近出现的SARS-2。冠状病毒E蛋白是一种小的、疏水的整体膜蛋白，在感染过程中寡聚形成通道，运输生理上重要的离子。这些蛋白是多功能的，在病毒的组装和发病过程中起着重要的作用。该奖项支持的研究将使用膜蛋白结构研究的尖端技术方法，包括异种表达系统、脂质立方相结晶（LCP）和时间分辨飞秒晶体学（TR-SFX）。在LCP中结晶的基本原理是，脂质提供了一个更类似于天然的环境，可以促进病毒孔蛋白正常低聚状态的重建，反过来，形成良好衍射的纳米晶体，可以使用x射线自由电子激光（XFEL）能力进行分析。该RAPID奖由生物基础设施部使用《冠状病毒援助、救济和经济安全法案》的资金颁发。该奖项反映了美国国家科学基金会的法定使命，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。

There is an urgent need to determine structures for Severe Acute Respiratory Syndrome Coronavirus (CoV)-2 (SARS-CoV-2) proteins. CoVs are enveloped viruses, assembled from four major structural proteins: membrane (M), spike (S), and envelope (E) and nucleocapsid (N). Researchers supported by this award will focus on the CoV E proteins to understand how the proteins facilitate, through their interactions with cellular membranes, the budding process as the viral genome is enveloped. The studies will facilitate identification and design of molecules for use in structure-function studies and may structural information that is critical for the design of therapeutics. Postdoctoral and graduate student trainees, including members from under-represented groups, will be integrally involved in the research effort. Results from these studies will be published in peer-reviewed journals, presented at scientific meetings, and made available through shared data repositories.The goal of the research supported by this award is to solve atomic structures of CoV E proteins from highly pathogenic human CoVs: Middle East respiratory syndrome (MERS), severe acute respiratory syndrome (SARS) and the recently emerged SARS-2. CoV E proteins are viroporins - small, hydrophobic integral membrane proteins that oligomerize to form channels that transport physiologically important ions during infection. The proteins are multifunctional, in that they play important roles in virus assembly and pathogenesis. Studies supported by this award will use of cutting-edge technology approaches for membrane protein structural studies, including heterologous expression systems, crystallization in lipid cubic phase (LCP), and time resolved femtosecond crystallography (TR-SFX). The rationale to crystalize in LCP is that the lipids provide a more native-like environment that may facilitate reconstitution of normal oligomeric states of viroporins and in turn, formation of well-diffracting nanocrystals that can be analyzed using X-ray free electron laser (XFEL) capabilities. This RAPID award is made by the Division of Biological Infrastructure 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.