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TitleStabilized coronavirus spikes are resistant to conformational changes induced by receptor recognition or proteolysis.
Journal, issue, pagesSci Rep, Vol. 8, Issue 1, Page 15701, Year 2018
Publish dateOct 24, 2018
AuthorsRobert N Kirchdoerfer / Nianshuang Wang / Jesper Pallesen / Daniel Wrapp / Hannah L Turner / Christopher A Cottrell / Kizzmekia S Corbett / Barney S Graham / Jason S McLellan / Andrew B Ward /
PubMed AbstractSevere acute respiratory syndrome coronavirus (SARS-CoV) emerged in 2002 as a highly transmissible pathogenic human betacoronavirus. The viral spike glycoprotein (S) utilizes angiotensin-converting ...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'.
External linksSci Rep / PubMed:30356097 / PubMed Central
MethodsEM (single particle)
Resolution3.2 - 13.2 Å
Structure data

EMDB-7573, PDB-6crv:
SARS Spike Glycoprotein, Stabilized variant, C3 symmetry
Method: EM (single particle) / Resolution: 3.2 Å

EMDB-7574, PDB-6crw:
SARS Spike Glycoprotein, Stabilized variant, single upwards S1 CTD conformation
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-7575, PDB-6crx:
SARS Spike Glycoprotein, Stabilized variant, two S1 CTDs in the upwards conformation
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-7576:
SARS spike glycoprotein, stabilized variant, three S1 CTDs in an upwards conformation
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-7577, PDB-6crz:
SARS Spike Glycoprotein, Trypsin-cleaved, Stabilized variant, C3 symmetry
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-7578, PDB-6cs0:
SARS Spike Glycoprotein, Trypsin-cleaved, Stabilized variant, one S1 CTD in an upwards conformation
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-7579, PDB-6cs1:
SARS Spike Glycoprotein, Trypsin-cleaved, Stabilized variant, two S1 CTDs in an upwards conformation
Method: EM (single particle) / Resolution: 4.6 Å

EMDB-7580:
SARS spike glycoprotein, trypsin-cleaved, stabilized variant, three S1 CTDs in an upwards conformation
Method: EM (single particle) / Resolution: 10.6 Å

EMDB-7581:
SARS spike glycoprotein, trypsin-cleaved, stabilized variant, three S1 CTDs in a downwards conformation
Method: EM (single particle) / Resolution: 5.3 Å

EMDB-7582, PDB-6cs2:
SARS Spike Glycoprotein - human ACE2 complex, Stabilized variant, all ACE2-bound particles
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-7584:
SARS spike glycoprotein, stabilized variant, ACE2-bound dataset, S1 CTD configuration: upwards, downwards, downwards
Method: EM (single particle) / Resolution: 6.6 Å

EMDB-7585:
SARS spike glycoprotein, stabilized variant, ACE2-bound dataset, S1 CTD configuration: upwards, upwards, downwards
Method: EM (single particle) / Resolution: 4.4 Å

EMDB-7586:
SARS spike glycoprotein, stabilized variant, S1 CTD configuration: ACE2-bound, downwards, downwards
Method: EM (single particle) / Resolution: 6.2 Å

EMDB-7601:
SARS spike glycoprotein, stabilized variant, S1 CTD configuration: ACE2-bound, downwards, upwards
Method: EM (single particle) / Resolution: 7.6 Å

EMDB-7602:
SARS spike glycoprotein, stabilized variant, S1 CTD configuration: ACE2-bound, upwards, downwards
Method: EM (single particle) / Resolution: 5.5 Å

EMDB-7603:
SARS spike glycoprotein, stabilized variant, S1 CTD configurations: ACE2-bound, upwards, upwards
Method: EM (single particle) / Resolution: 13.2 Å

EMDB-7604:
SARS spike glycoprotein, stabilized variant, S1 CTD configurations: ACE2-bound, ACE2-bound, downwards
Method: EM (single particle) / Resolution: 8.1 Å

EMDB-7605:
SARS spike glycoprotein, stabilized variant, S1 CTD configurations: ACE2-bound, ACE2-bound, upwards
Method: EM (single particle) / Resolution: 8.8 Å

EMDB-7606:
SARS spike glycoprotein, stabilized variant, S1 CTD configurations: ACE2-bound, ACE2-bound, ACE2-bound
Method: EM (single particle) / Resolution: 9.3 Å

EMDB-7607:
SARS spike glycoprotein, stabilized variant, S1 CTD configurations: ACE2-bound, upwards/downwards mix, downwards
Method: EM (single particle) / Resolution: 4.6 Å

EMDB-7608:
SARS spike glycoprotein, focused refinement of S1 CTD bound to ACE2
Method: EM (single particle) / Resolution: 7.9 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose

Source
  • SARS coronavirus
  • enterobacteria phage t4 (virus)
  • homo sapiens (human)
  • human sars coronavirus
KeywordsVIRAL PROTEIN / membrane fusion / glycoprotein / receptor binding / viral protein/hydrolase / viral protein-hydrolase complex

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