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Title:Cryo-electron microscopy reconstruction of the Enterovirus 71 empty capsid
Authors:Shingler KL, Yoder JL, Carnegie MS, Ashley RE, Makhov AM, Conway JF, Hafenstein S
Sample:Enterovirus 71 empty capsid
Method:Single particle reconstruction (9.2 angstroms resolution)
Red flagLatest update:2013-04-17
Sample
Sample name: Enterovirus 71 empty capsid
Oligomeric state: icosahedral virus
Theoretical molecular weight of the sample: 7
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusHuman enterovirus 717Human enterovirus 71purified virus in solution
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.50.1 mg/mL10 mM Tris-HCl, 20 mM NaCl, 5 mM MgCl2glow discharged holey carbon Quantifoil electron microscopy grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE-PROPANE MIXTURE95%77 KFEI VITROBOT MARK III ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2 mm1730 nm4370 nm5000050000FIELD EMISSION GUNKODAK SO-163 FILM mmCTFFIND3 (EMAN)

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°° eV95 K K K mrad15 e/Å217-APR-2012
Processing
Protocol:Common Lines
Software:EMAN2, auto3DEM
CTF correction:auto3DEM
Number of particles:1931
Imposed symmetry:I
Resolution by author:9.2 Å
Resolution method:FSC 0.5
Processing details:A single data set was used for the reconstruction. These particles were selected individually.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
406.35 μm/pixelNIKON SUPER COOLSCAN 9000
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
3VBU rigid bodycorrelation coefficientChimera, SitusREALProtocol: rigid body. The protomer was fit in Chimera, used to generate a full crystal map, then docked as a rigid body in Situs.