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- EMDB-11497: Cryo-EM structure of SARS-CoV-2 Spike Proteins on intact virions:... -

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Basic information

Entry
Database: EMDB / ID: EMD-11497
TitleCryo-EM structure of SARS-CoV-2 Spike Proteins on intact virions: 3 Closed RBDs
Map data
SampleSevere acute respiratory syndrome coronavirus 2:
virus / Spike glycoprotein / ligand
Function / homology
Function and homology information


host cell endoplasmic reticulum-Golgi intermediate compartment membrane / receptor-mediated virion attachment to host cell / host cell surface receptor binding / viral entry into host cell / fusion of virus membrane with host endosome membrane / viral envelope / pathogenesis / host cell plasma membrane / virion membrane / integral component of membrane
Spike glycoprotein S2, coronavirus / Spike receptor binding domain, betacoronavirus / Spike glycoprotein, heptad repeat 2, coronavirus / Spike receptor binding domain superfamily, coronavirus / Spike glycoprotein S2 superfamily, coronavirus
Spike glycoprotein
Biological speciesSevere acute respiratory syndrome coronavirus 2 / 2019-nCoV (virus)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.5 Å
AuthorsKe Z / Qu K / Nakane T / Xiong X / Cortese M / Zila V / Scheres SHW / Briggs JAG
Funding supportEuropean Union, United Kingdom, Japan, Germany, 5 items
OrganizationGrant numberCountry
European Research Council (ERC)ERC-CoG-648432European Union
Medical Research Council (MRC, United Kingdom)MC_UP_A025_1013 United Kingdom
Medical Research Council (MRC, United Kingdom)MC_UP_1201/16 United Kingdom
Japan Society for the Promotion of Science (JSPS) Japan
German Research Foundation (DFG)240245660-SFB 1129 Germany
CitationJournal: Nature / Year: 2020
Title: Structures and distributions of SARS-CoV-2 spike proteins on intact virions.
Authors: Zunlong Ke / Joaquin Oton / Kun Qu / Mirko Cortese / Vojtech Zila / Lesley McKeane / Takanori Nakane / Jasenko Zivanov / Christopher J Neufeldt / Berati Cerikan / John M Lu / Julia Peukes / ...Authors: Zunlong Ke / Joaquin Oton / Kun Qu / Mirko Cortese / Vojtech Zila / Lesley McKeane / Takanori Nakane / Jasenko Zivanov / Christopher J Neufeldt / Berati Cerikan / John M Lu / Julia Peukes / Xiaoli Xiong / Hans-Georg Kräusslich / Sjors H W Scheres / Ralf Bartenschlager / John A G Briggs /
Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions are surrounded by a lipid bilayer from which spike (S) protein trimers protrude. Heavily glycosylated S trimers bind the ACE2 ...Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virions are surrounded by a lipid bilayer from which spike (S) protein trimers protrude. Heavily glycosylated S trimers bind the ACE2 receptor and mediate entry of virions into target cells. S exhibits extensive conformational flexibility: it modulates exposure of its receptor binding site and later undergoes complete structural rearrangement to drive fusion of viral and cellular membranes. The structures and conformations of soluble, overexpressed, purified S proteins have been studied in detail using cryo-electron microscopy. The structure and distribution of S on the virion surface, however, has not been characterized. Here we applied cryo-electron microscopy and tomography to image intact SARS-CoV-2 virions, determining the high-resolution structure, conformational flexibility and distribution of S trimers in situ on the virion surface. These results reveal the conformations of S present on the virion, and provide a basis from which to understand interactions between S and neutralizing antibodies during infection or vaccination.
Validation ReportPDB-ID: 6zwv

SummaryFull reportAbout validation report
History
DepositionJul 28, 2020-
Header (metadata) releaseAug 5, 2020-
Map releaseAug 5, 2020-
UpdateSep 2, 2020-
Current statusSep 2, 2020Processing site: PDBe / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.0261
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by cylindrical radius
  • Surface level: 0.0261
  • Imaged by UCSF Chimera
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  • Surface view with fitted model
  • Atomic models: PDB-6zwv
  • Surface level: 0.0261
  • Imaged by UCSF Chimera
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Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

Downloads & links

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Map

FileDownload / File: emd_11497.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.06 Å/pix.
x 256 pix.
= 271.616 Å
1.06 Å/pix.
x 256 pix.
= 271.616 Å
1.06 Å/pix.
x 256 pix.
= 271.616 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.061 Å
Density
Contour LevelBy AUTHOR: 0.0261 / Movie #1: 0.0261
Minimum - Maximum-0.065178536 - 0.15941393
Average (Standard dev.)0.00023489696 (±0.00522143)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions256256256
Spacing256256256
CellA=B=C: 271.616 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.0611.0611.061
M x/y/z256256256
origin x/y/z0.0000.0000.000
length x/y/z271.616271.616271.616
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS256256256
D min/max/mean-0.0650.1590.000

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Supplemental data

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Half map: #2

Fileemd_11497_half_map_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Half map: #1

Fileemd_11497_half_map_2.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

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Sample components

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Entire Severe acute respiratory syndrome coronavirus 2

EntireName: Severe acute respiratory syndrome coronavirus 2
Details: sucrose-cushion purified SARS-CoV-2 virions produced from infected VeroE6 cells.
Number of components: 3

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Component #1: virus, Severe acute respiratory syndrome coronavirus 2

VirusName: Severe acute respiratory syndrome coronavirus 2 / Class: VIRION
Details: sucrose-cushion purified SARS-CoV-2 virions produced from infected VeroE6 cells.
Empty: No / Enveloped: Yes / Isolate: STRAIN
SpeciesSpecies: Severe acute respiratory syndrome coronavirus 2 / Strain: Germany/BavPat1/2020

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Component #2: protein, Spike glycoprotein

ProteinName: Spike glycoprotein / Number of Copies: 3 / Recombinant expression: No
MassTheoretical: 141.239391 kDa
SourceSpecies: 2019-nCoV (virus)

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Component #3: ligand, 2-acetamido-2-deoxy-beta-D-glucopyranose

LigandName: 2-acetamido-2-deoxy-beta-D-glucopyranose / Number of Copies: 33 / Recombinant expression: No
MassTheoretical: 0.221208 kDa

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Experimental details

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Sample preparation

SpecimenSpecimen state: Particle / Method: cryo EM
Sample solutionpH: 7.4
VitrificationInstrument: LEICA EM GP / Cryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
ImagingMicroscope: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 50 e/Å2 / Illumination mode: FLOOD BEAM
LensMagnification: 81000 X (nominal) / Cs: 2.7 mm / Imaging mode: BRIGHT FIELD / Defocus: 1000.0 - 3000.0 nm / Energy filter: GIF Bioquantum
Specimen HolderModel: FEI TITAN KRIOS AUTOGRID HOLDER
CameraDetector: OTHER

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Image acquisition

Image acquisitionNumber of digital images: 7982 / Sampling size: 5 µm

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Image processing

ProcessingMethod: single particle reconstruction / Applied symmetry: C3 (3 fold cyclic) / Number of projections: 29183
3D reconstructionSoftware: RELION / Resolution: 3.5 Å / Resolution method: FSC 0.143 CUT-OFF

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Atomic model buiding

Modeling #1Refinement protocol: flexible
Input PDB model: 6ZP0
Output model

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