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Title:Cryo-electron tomography of glycoprotein lacking the mucin-like domain 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 glycoprotein trimer lacking the mucin-like domain
Method:Subtomogram averaging
Red flagLatest update:2014-09-03
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Sample
Sample name: Molecular structure of Ebola VLP glycoprotein trimer lacking the mucin-like domain
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 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 IVBlot for 4 seconds at 22 degrees C, 100% humidity, blot force 4, 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/Å216-MAY-2013
Processing
Software:IMOD
Number of subtomogram averaged:6304
Imposed symmetry:C3
Processing details:Subtomogram density was selected using an automatic selection program.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
29 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
3CSY rigid bodyChimeraREALProtocol: rigid body, automated fitting procedures