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- PDB-9r6s: Prefusion stabilized spike glycoprotein of human coronavirus OC43. -

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Basic information

Entry
Database: PDB / ID: 9r6s
TitlePrefusion stabilized spike glycoprotein of human coronavirus OC43.
ComponentsSpike glycoprotein
KeywordsVIRAL PROTEIN / fusion glycoprotein / membrane protein
Function / homology
Function and homology information


host cell endoplasmic reticulum-Golgi intermediate compartment membrane / receptor-mediated virion attachment to host cell / endocytosis involved in viral entry into host cell / fusion of virus membrane with host plasma membrane / fusion of virus membrane with host endosome membrane / viral envelope / host cell plasma membrane / virion membrane / membrane
Similarity search - Function
Spike (S) protein S1 subunit, receptor-binding domain, HCoV-OC43-like / Spike (S) protein S1 subunit, N-terminal domain, murine hepatitis virus-like / Coronavirus spike glycoprotein S1, C-terminal / Coronavirus spike glycoprotein S1, C-terminal / Spike glycoprotein, N-terminal domain superfamily / Spike S1 subunit, receptor binding domain superfamily, betacoronavirus / Spike glycoprotein, betacoronavirus / Betacoronavirus spike (S) glycoprotein S1 subunit N-terminal (NTD) domain profile. / Spike glycoprotein S1, N-terminal domain, betacoronavirus-like / Betacoronavirus-like spike glycoprotein S1, N-terminal ...Spike (S) protein S1 subunit, receptor-binding domain, HCoV-OC43-like / Spike (S) protein S1 subunit, N-terminal domain, murine hepatitis virus-like / Coronavirus spike glycoprotein S1, C-terminal / Coronavirus spike glycoprotein S1, C-terminal / Spike glycoprotein, N-terminal domain superfamily / Spike S1 subunit, receptor binding domain superfamily, betacoronavirus / Spike glycoprotein, betacoronavirus / Betacoronavirus spike (S) glycoprotein S1 subunit N-terminal (NTD) domain profile. / Spike glycoprotein S1, N-terminal domain, betacoronavirus-like / Betacoronavirus-like spike glycoprotein S1, N-terminal / Betacoronavirus spike (S) glycoprotein S1 subunit C-terminal (CTD) domain profile. / Spike (S) protein S1 subunit, receptor-binding domain, betacoronavirus / Betacoronavirus spike glycoprotein S1, receptor binding / Spike glycoprotein S2 superfamily, coronavirus / Spike glycoprotein S2, coronavirus, heptad repeat 1 / Spike glycoprotein S2, coronavirus, heptad repeat 2 / Coronavirus spike (S) glycoprotein S2 subunit heptad repeat 1 (HR1) region profile. / Coronavirus spike (S) glycoprotein S2 subunit heptad repeat 2 (HR2) region profile. / Spike glycoprotein S2, coronavirus / Coronavirus spike glycoprotein S2
Similarity search - Domain/homology
Sapienic acid / Spike glycoprotein
Similarity search - Component
Biological speciesHuman coronavirus OC43
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.1 Å
AuthorsMelchers, J.M. / Hulswit, R.J.G. / Hurdiss, D.L.
Funding support Netherlands, 1items
OrganizationGrant numberCountry
Other private Netherlands
CitationJournal: To Be Published
Title: AI-guided prefusion stabilization of the human coronavirus OC43 spike protein enables universal Embecovirus antigen design.
Authors: Melchers, J.M. / Juraszek, J. / Hulswit, R.J.G. / van Overveld, D. / Le, L. / van Kuppeveld, F. / Hurdiss, D.L. / Bosch, B.J. / Langedijk, J.P.M. / Bakkers, M.J.G.
History
DepositionMay 13, 2025Deposition site: PDBE / Processing site: PDBE
Revision 1.0Mar 11, 2026Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: EM metadata / Data content type: EM metadata / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: FSC / Data content type: FSC / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: Half map / Part number: 1 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: Half map / Part number: 2 / Data content type: Half map / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: Image / Data content type: Image / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: Mask / Part number: 1 / Data content type: Mask / Provider: repository / Type: Initial release
Revision 1.0Mar 11, 2026Data content type: Primary map / Data content type: Primary map / Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

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Assembly

Deposited unit
A: Spike glycoprotein
B: Spike glycoprotein
C: Spike glycoprotein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)447,73057
Polymers432,0283
Non-polymers15,70254
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: electron microscopy, not applicable
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
d_1ens_1chain "C"
d_2ens_1chain "B"
d_3ens_1chain "A"

NCS domain segments:

Ens-ID: ens_1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
d_11VALVALASNASNCC15 - 122915 - 1229
d_12NAGNAGNAGNAGCUA1302
d_13NAGNAGNAGNAGCVA1303
d_14NAGNAGNAGNAGPP1
d_15NAGNAGNAGNAGQQ1
d_16NAGNAGNAGNAGQQ2
d_17NAGNAGNAGNAGPP2
d_18NAGNAGNAGNAGCWA1304
d_19NAGNAGNAGNAGCXA1305
d_110NAGNAGNAGNAGRR1
d_111NAGNAGNAGNAGRR2
d_112NAGNAGNAGNAGCYA1306
d_113NAGNAGNAGNAGSS1
d_114NAGNAGNAGNAGSS2
d_115NAGNAGNAGNAGTT1
d_116NAGNAGNAGNAGTT2
d_117NAGNAGNAGNAGCZA1307
d_118NAGNAGNAGNAGCAB1308
d_119NAGNAGNAGNAGCBB1309
d_120NAGNAGNAGNAGCCB1310
d_121NAGNAGNAGNAGUU1
d_122NAGNAGNAGNAGUU2
d_1238Z98Z98Z98Z9BSA1312
d_124NAGNAGNAGNAGCDB1311
d_125NAGNAGNAGNAGCEB1312
d_21VALVALASNASNBB15 - 122915 - 1229
d_22NAGNAGNAGNAGBHA1301
d_23NAGNAGNAGNAGBIA1302
d_24NAGNAGNAGNAGJJ1
d_25NAGNAGNAGNAGKK1
d_26NAGNAGNAGNAGKK2
d_27NAGNAGNAGNAGJJ2
d_28NAGNAGNAGNAGBJA1303
d_29NAGNAGNAGNAGBKA1304
d_210NAGNAGNAGNAGLL1
d_211NAGNAGNAGNAGLL2
d_212NAGNAGNAGNAGBLA1305
d_213NAGNAGNAGNAGMM1
d_214NAGNAGNAGNAGMM2
d_215NAGNAGNAGNAGNN1
d_216NAGNAGNAGNAGNN2
d_217NAGNAGNAGNAGBMA1306
d_218NAGNAGNAGNAGBNA1307
d_219NAGNAGNAGNAGBOA1308
d_220NAGNAGNAGNAGBPA1309
d_221NAGNAGNAGNAGOO1
d_222NAGNAGNAGNAGOO2
d_2238Z98Z98Z98Z9AGA1312
d_224NAGNAGNAGNAGBQA1310
d_225NAGNAGNAGNAGBRA1311
d_31VALVALASNASNAA15 - 122915 - 1229
d_32NAGNAGNAGNAGAV1301
d_33NAGNAGNAGNAGAW1302
d_34NAGNAGNAGNAGDD1
d_35NAGNAGNAGNAGEE1
d_36NAGNAGNAGNAGEE2
d_37NAGNAGNAGNAGDD2
d_38NAGNAGNAGNAGAX1303
d_39NAGNAGNAGNAGAY1304
d_310NAGNAGNAGNAGFF1
d_311NAGNAGNAGNAGFF2
d_312NAGNAGNAGNAGAZ1305
d_313NAGNAGNAGNAGGG1
d_314NAGNAGNAGNAGGG2
d_315NAGNAGNAGNAGHH1
d_316NAGNAGNAGNAGHH2
d_317NAGNAGNAGNAGAAA1306
d_318NAGNAGNAGNAGABA1307
d_319NAGNAGNAGNAGACA1308
d_320NAGNAGNAGNAGADA1309
d_321NAGNAGNAGNAGII1
d_322NAGNAGNAGNAGII2
d_3238Z98Z98Z98Z9CTA1301
d_324NAGNAGNAGNAGAEA1310
d_325NAGNAGNAGNAGAFA1311

NCS oper:
IDCodeMatrixVector
1given(-0.499985563361, 0.866033737499, 4.40993794767E-5), (-0.86603373861, -0.499985562448, -3.05329156729E-5), (-4.39348202566E-6, -5.34575675224E-5, 0.999999998561)132.484094043, 494.506849995, 0.0161293412641
2given(-0.500132488484, -0.865948890145, 0.0001167021337), (0.865948884453, -0.500132499959, -0.000109536576206), (0.000153219606474, 4.62752820533E-5, 0.999999987191)494.494216428, 132.580381046, -0.0331883759127

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Components

#1: Protein Spike glycoprotein / S glycoprotein / E2 / Peplomer protein


Mass: 144009.203 Da / Num. of mol.: 3
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Human coronavirus OC43 / Cell line (production host): Expi293 / Production host: Homo sapiens (human) / References: UniProt: A0A3S6FHI0
#2: Polysaccharide
2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose


Type: oligosaccharide / Mass: 424.401 Da / Num. of mol.: 18
Source method: isolated from a genetically manipulated source
DescriptorTypeProgram
DGlcpNAcb1-4DGlcpNAcb1-ROHGlycam Condensed SequenceGMML 1.0
WURCS=2.0/1,2,1/[a2122h-1b_1-5_2*NCC/3=O]/1-1/a4-b1WURCSPDB2Glycan 1.1.0
[][D-1-deoxy-GlcpNAc]{[(4+1)][b-D-GlcpNAc]{}}LINUCSPDB-CARE
#3: Sugar...
ChemComp-NAG / 2-acetamido-2-deoxy-beta-D-glucopyranose / N-acetyl-beta-D-glucosamine / 2-acetamido-2-deoxy-beta-D-glucose / 2-acetamido-2-deoxy-D-glucose / 2-acetamido-2-deoxy-glucose / N-ACETYL-D-GLUCOSAMINE


Type: D-saccharide, beta linking / Mass: 221.208 Da / Num. of mol.: 33 / Source method: obtained synthetically / Formula: C8H15NO6
IdentifierTypeProgram
DGlcpNAcbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-b-D-glucopyranosamineCOMMON NAMEGMML 1.0
b-D-GlcpNAcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcNAcSNFG CARBOHYDRATE SYMBOLGMML 1.0
#4: Chemical ChemComp-8Z9 / Sapienic acid


Mass: 254.408 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C16H30O2 / Feature type: SUBJECT OF INVESTIGATION
Has ligand of interestY
Has protein modificationY

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Trimeric spike glycoprotein / Type: COMPLEX / Entity ID: #1 / Source: RECOMBINANT
Molecular weightValue: 0.452 MDa / Experimental value: YES
Source (natural)Organism: Human coronavirus OC43 / Strain: ANZ78834.1 / Cellular location: membrane
Source (recombinant)Organism: Homo sapiens (human) / Cell: Expi293
Buffer solutionpH: 7.4 / Details: 20 mM Tris-HCL, 150 mM NaCl, pH 7.4
Buffer component
IDConc.NameFormulaBuffer-ID
120 mMTris1
2150 mMsodium chlorideNaCl1
SpecimenConc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen supportGrid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R1.2/1.3
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 298 K

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: TFS KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD / Nominal magnification: 105000 X / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm
Specimen holderSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Image recordingElectron dose: 50 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k)
EM imaging opticsEnergyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV

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Processing

EM software
IDNameVersionCategory
1cryoSPARCparticle selection
2EPUimage acquisition
4cryoSPARCCTF correction
7UCSF ChimeraX1.9model fitting
8ISOLDEmodel fitting
10cryoSPARCinitial Euler assignment
11cryoSPARCfinal Euler assignment
13cryoSPARC3D reconstruction
14PHENIX1.20.1_4487:model refinement
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
SymmetryPoint symmetry: C3 (3 fold cyclic)
3D reconstructionResolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 92153 / Symmetry type: POINT
Atomic model building
IDPDB-ID 3D fitting-IDAccession codeInitial refinement model-IDSource nameType
16NZK16NZK1PDBexperimental model
21AlphaFoldin silico model
RefinementCross valid method: NONE
Stereochemistry target values: GeoStd + Monomer Library + CDL v1.2
Displacement parametersBiso mean: 32.33 Å2
Refine LS restraints
Refine-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.001729088
ELECTRON MICROSCOPYf_angle_d0.449139576
ELECTRON MICROSCOPYf_chiral_restr0.0414638
ELECTRON MICROSCOPYf_plane_restr0.00375022
ELECTRON MICROSCOPYf_dihedral_angle_d4.57884254
Refine LS restraints NCS
Ens-IDDom-IDAsym-IDAuth asym-IDRefine-IDTypeRms dev position (Å)
ens_1d_2CCELECTRON MICROSCOPYNCS constraints1.11752181924E-10
ens_1d_3CCELECTRON MICROSCOPYNCS constraints2.23560831186E-12

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