Stabilized coronavirus spikes are resistant to conformational changes induced by receptor recognition or proteolysis

By R. Kirchdoerfer, Nianshuang Wang, J. Pallesen, D. Wrapp, H. Turner, C. Cottrell, Kizzmekia S. Corbett, Barney S. Graham, J. McLellan, A. Ward. 2018

479 citations recorded by the source corpus.

Severe acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as a highly transmissible pathogenic human betacoronavirus. The viral spike glycoprotein (S) utilizes angiotensin-converting enzyme 2 (ACE2) as a host protein receptor and mediates fusion of the viral and host membranes, making S essential to viral entry into host cells and host species tropism. As SARS-CoV enters host cells, the viral S is believed to undergo a number of conformational transitions as it is cleaved by host proteases and binds to host receptors. We recently developed stabilizing mutations for coronavirus spikes that prevent the transition from the pre-fusion to post-fusion states. Here, we present cryo-EM analyses of a stabilized trimeric SARS-CoV S, as well as the trypsin-cleaved, stabilized S, and its interactions with ACE2. Neither binding to ACE2 nor cleavage by trypsin at the S1/S2 cleavage site impart large conformational changes within stabilized SARS-CoV S or expose the secondary cleavage site, S2′.

Subfields: Animal Science and Zoology, Infectious Diseases

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Citations in the corpus

Elicitation of Potent Neutralizing Antibody Responses by Designed Protein Nanoparticle Vaccines for SARS-CoV-2 (2020, neutral): “…purified RBD-I53-50A proteins (~30 mg purified protein per liter Expi293F cells) were ~20-fold higher than for the prefusion-stabilized S-2P trimer (Kirchdoerfer et al., 2018; Pallesen et al., 2017; Walls et al., 2020; Wrapp et al., 2020) (~1.5 mg/L), and increased to ~60 mg/L following promoter…”
The Molecular Docking Study of Potential Drug Candidates Showing Anti-COVID-19 Activity by Exploring of Therapeutic Targets of SARS-CoV-2 (2020, background): “SARS Spike glycoprotein-Human ACE2 complex[17] 6CS2 CI”
Structure, Function, and Antigenicity of the SARS-CoV-2 Spike Glycoprotein (2020, methodology): “As is the case for other b-coronavirus S glycoproteins, including SARS-CoV S (Gui et al., 2017; Kirchdoerfer et al., 2018; Pallesen et al., 2017; Park et al., 2019; Song et al., 2018; Tortorici et al., 2019;Walls et al., 2016a;Walls et al., 2016b;Walls et al., 2019; Yuan et al., 2017), the S1 subuni”
A highly conserved cryptic epitope in the receptor-binding domains of SARS-CoV-2 and SARS-CoV (2020, background): “protein is mostly found in the none-“up”, single-“up”, or double-”up” conformations (19, 122 21), but rarely in the triple-“up” conformation, even with ACE2 receptor bound (21, 22).”
Cryo-EM Structure of the 2019-nCoV Spike in the Prefusion Conformation (2020, methodology): “proline mutations in the C-terminal S2 fusion machinery based on a previous stabilization 39 strategy which proved highly effective for betacoronavirus S proteins (11, 14).”
Structure-guided covalent stabilization of coronavirus spike glycoprotein trimers in the closed conformation (2020, unknown): “Finally, the introduction of double proline substitutions, herein 2P, to prevent fusogenic conformational changes of MERS-CoV S 20 and SARS-CoV S 55 was shown to stabilize the prefusion states of these glycoproteins.”
Controlling the SARS-CoV-2 spike glycoprotein conformation (2020, unknown): “In the u1S2q data, we observed an S-protein population with 2 RBDs in the ‘up’ position which has been reported before for the SARS-CoV-1 and MERS CoV spike ectodomains 7,12 .”
The human coronavirus HCoV-229E S-protein structure and receptor binding (2019, background): “741 Two views showing a morphing between the two S-proteins.”