RAPID: IIBR: Coronavirus Viroporin XFEL Structural Studies

Arizona State University, Scottsdale AZ

Investigators

Abstract

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.