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Title:Structure of furin-cleaved immature dengue virus at low pH
Authors:I.Yu,H.A.Holdaway,P.R.Chipman,R.J.Kuhn,M.G.Rossmann,J.Chen
Sample:Immature dengue virus at low pH after furin cleavage
Aggregation state:Single particle (22 angstroms resolution)
Red flagLatest update:2011-09-27
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Sample
Sample name: Immature dengue virus at low pH after furin cleavage
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1virusDengue virusDengue virus00.026.0.01.013
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
6 mg/mL6 mM Tris-HCL, 25 mM MES, 120 mM NaCl, 1 mM EDTA
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KNONEA small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by the means of fine tweezers. Once the ethane in the vial is completely frozen, it needs to be slightly melted. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot of excess buffer, sufficient to leave a thin layer on the grid. After a predetermined time, the filter paper is removed, and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen, and the grid is transferred under liquid nitrogen to a storage box immersed liquid nitrogen for later use in the microscope. ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG200 kVFLOOD BEAMBRIGHT FIELD mm1800 nm3800 nm50000FIELD EMISSION GUNGatan MSC (multiscan) 4000 (4k x 4k) mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
EucentricGATAN LIQUID NITROGEN°° eV K K K mrad25 e/Å213-JUN-2008
Processing
Software:EM3DR
CTF correction:Each particle
Resolution by author:22 Å
Resolution method:FSC at 0.5 cut-off
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
45 μm/pixellink
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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