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Title:Electron cryo-microscopy of escrt-III helical polymer
Authors:Effantin G, Dordor A, Sandrin V, Martinelli N, Sundquist WI, Schoehn G, Weissenhorn W
Sample:ESCRT-III Chmp2a - Chmp3 co-polymer
Method:Helical reconstruction (22.4 angstroms resolution)
Red flagLatest update:2013-01-23
Sample
Sample name: ESCRT-III Chmp2a - Chmp3 co-polymer
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
1proteinchmp2atrueHomo sapienschmp2a mbp tag was cleaved by tev protease before imaging
2proteinchmp3trueHomo sapiens
Experiment
Specimen state: Filament
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.6 mg/mL20 mM Hepes pH 7.6, 150 mM NaCl400 mesh copper quantifoil grid
Helical parameters:
HandPhi incrementAzimuthal (Z) incrementRotational symmetry
RIGHT HANDED11.9°6.03 Åc6
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100% KFEI VITROBOT MARK II ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F30300 kVSPOT SCANBRIGHT FIELD2.3 mm nm nm39000FIELD EMISSION GUNKODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
SIDE ENTRY, EUCENTRIC°° eV K K K mrad15 e/Å210-OCT-2010
Processing
Protocol:ihrsr
Software:ctffind, bsof, spider, imagic
CTF correction:each particle
Resolution by author:22.4 Å
Resolution method:FSC 0.5
Processing details:The particles were aligned using IHRSR.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
787 μm/pixel8ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails