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Title:Cryo-electron tomography of full-length glycoprotein from Ebola virus-like particles
Authors:Tran EEH, Simmons JA, Bartesaghi A, Shoemaker CJ, Nelson E, White JM, Subramaniam S
Sample:Molecular structure of Ebola VLP full-length glycoprotein trimer
Method:Subtomogram averaging
Red flagLatest update:2014-09-03
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Sample
Sample name: Molecular structure of Ebola VLP full-length glycoprotein trimer
Oligomeric state: trimer
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinEnvelope glycoproteintrimertrueZaire ebolavirusEnvelope glycoproteins present on the surface of intact virus-like particles
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.4 mg/mL130 mM NaCl, 20 mM HEPES, 10% sucrose200 mesh Quantifoil Multi-A
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%77 KFEI VITROBOT MARK IIIBlot for 6 seconds at 22 degrees C, 100% humidity, blot offset -2, plunge into an ethane slurry cooled by liquid nitrogen. ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300200 kVFLOOD BEAMBRIGHT FIELD mm2500 nm2500 nm34000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN-60°60°GATAN GIF0-20 eV81 K K K mrad150 e/Å205-MAR-2013
Processing
Software:IMOD
Number of subtomogram averaged:5298
Imposed symmetry:C3
Processing details:Subtomogram density was selected using an automatic selection program.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
37 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
3CSY rigid bodyChimeraREALProtocol: rigid body, automated fitting procedures