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Title:Structure of herpesvirus fusion glycoprotein B-bilayer complex revealing the protein-membrane and lateral protein-protein interaction
Authors:Maurer UE, Zeev-Ben-Mordehai Z, Pandurangan AP, Cairns TM, Hannah BP, Whitbeck JC, Eisenberg RJ, Cohen GH, Topf M, Huiskonen JT, Grunewald K
Sample:HSV-1 glycoprotein B ectodomain lacking the membrane-proximal region bound to a lipid bilayer
Method:Subtomogram averaging (27 angstroms resolution)
Red flagLatest update:2013-08-21
Sample
Sample name: HSV-1 glycoprotein B ectodomain lacking the membrane-proximal region bound to a lipid bilayer
Oligomeric state: One trimer of gB bound to a lipid bilayer
Experimental molecular weight of the sample: 0.259
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
1proteinEnvelope glycoprotein BTrimertrueHuman herpesvirus 1P06437GO:0046718IPR000234Liposomes consisting of phosphatidylcholine and cholesterol at 1.7:1 molar ratio were incubated with gB at pH 5.5 at 37C for one hour
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
5.51.0 mg/mLPBS with sodium citrateCflat
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE-PROPANE MIXTURE%120 KOTHERBlot manually for 3 s before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2 mm2000 nm2000 nm67000FIELD 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
liquid nitrogen cooledSIDE ENTRY, EUCENTRIC-60°60° eV K K K mrad100 e/Å217-JAN-2008
Processing
Protocol:Subtomogram averaging
Software:Jsubtomo
CTF correction:Low pass filter to the first zero crossing of the CTF
Number of subtomogram averaged:996
Imposed symmetry:C3
Resolution by author:27 Å
Resolution method:FSC 0.5
Processing details:The best 786 spikes (of 996) were selected based on constrained cross correlation coefficient and by excluding overlaps. All three Euler angles of the spike were refined. The sub-tomograms were picked manually from tomographic reconstructions of 38 liposomes.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
915 μm/pixel12The dataset consists of 9 tomograms (containing 38 liposomes with bound gB). Data were binned by factor of 2.
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
3NWF flexibleFlex-EMREAL