Abstract SARS-CoV-2 infection is controlled by the opening of the spike protein receptor binding domain (RBD), which transitions from a glycan-shielded (down) to an exposed (up) state in order to bind the human ACE2 receptor and infect cells. While snapshots of the up and down states have been obtained by cryoEM and cryoET, details of the RBD opening transition evade experimental characterization. Here, over 200 μs of weighted ensemble (WE) simulations of the fully glycosylated spike ectodomain allow us to characterize more than 300 continuous, kinetically unbiased RBD opening pathways. Together with biolayer interferometry experiments, we reveal a gating role for the N-glycan at position N343, which facilitates RBD opening. Residues D405, R408, and D427 also participate. The atomic-level characterization of the glycosylated spike activation mechanism provided herein achieves a new high-water mark for ensemble pathway simulations and offers a foundation for understanding the fundamental mechanisms of SARS-CoV-2 viral entry and infection. Competing Interest Statement The authors have declared no competing interest. Footnotes * ↵* contact authors: ramaro{at}ucsd.edu, ltchong{at}pitt.edu * https://amarolab.ucsd.edu/files/covid19/TRAJECTORIES_continuous_spike_opening_WE_chong_and_amarolab.tar.gz...

A glycan gate controls opening of the SARS-CoV-2 spike protein

10.1101/2021.02.15.431212