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Title:Cryo-electron microscopy reconstruction of the Enterovirus 71 A-particle
Authors:Shingler KL, Yoder JL, Carnegie MS, Ashley RE, Makhov AM, Conway JF, Hafenstein S
Sample:Enterovirus 71 A-Particle
Method:Single particle reconstruction (6.3 angstroms resolution)
Red flagLatest update:2013-04-17
Sample
Sample name: Enterovirus 71 A-Particle
Oligomeric state: icosahedral virus
Experimental molecular weight of the sample: 7
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1virusHuman enterovirus 7177Human 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%76 KFEI VITROBOT MARK III ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVSPOT SCANBRIGHT FIELD2 mm1730 nm4280 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:11436
Imposed symmetry:I
Resolution by author:6.3 Å
Resolution method:FSC 0.5
Processing details:A single data set was used for the reconstruction. The particles were selected using an automatic selection program (EMAN).
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. One protomer was fit in Chimera, used to generate a full crystal map, then docked as a rigid body using Situs into the cryoEM density map.