Controlling the SARS-CoV-2 Spike Glycoprotein Conformation

Henderson, Rory; Edwards, Robert J; Mansouri, Katayoun; Janowska, Katarzyna; Stalls, Victoria; Gobeil, Sophie; Kopp, Megan; Hsu, Allen; Borgnia, Mario; Parks, Rob; Haynes, Barton F.; Acharya, Priyamvada

The coronavirus (CoV) viral host cell fusion spike (S) protein is the primary immunogenic target for virus neutralization and the current focus of many vaccine design efforts. The highly flexible S-protein, with its mobile domains, presents a moving target to the immune system. Here, to better understand S-protein mobility, we implemented a structure-based vector analysis of available β-CoV S-protein structures. We found that despite overall similarity in domain organization, different β-CoV strains display distinct S-protein configurations. Based on this analysis, we developed two soluble ectodomain constructs in which the highly immunogenic and mobile receptor binding domain (RBD) is locked in either the all-RBDs ‘down’ position or is induced to display a previously unobserved in SARS-CoV-2 2-RBDs ‘up’ configuration. These results demonstrate that the conformation of the S-protein can be controlled via rational design and provide a framework for the development of engineered coronavirus spike proteins for vaccine applications.