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Title:Cryo EM reconstruction of mammalian 80S ribosome in classic PRE state 2
Authors:Budkevich T, Giesebrecht J, Altman R, Munro J, Mielke T, Nierhaus K, Blanchard S, Spahn C
Sample:80S-PRE-classic-2 complex from rabbit (Oryctolagus cuniculus) liver
Method:Single particle reconstruction (9 angstroms resolution)
Red flagLatest update:2013-03-06
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Sample
Sample name: 80S-PRE-classic-2 complex from rabbit (Oryctolagus cuniculus) liver
Oligomeric state: monomer
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1ribosome-eukaryote80S ribosomefalseOryctolagus cuniculus
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.5 mg/mLQuantifoil grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%77 KFEI VITROBOTFlash-frozen in liquid ethane on carbon coated Quantifoil grids msVitrification instrument: VitroBot (FEI)
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVSPOT SCANBRIGHT FIELD2 mm2000 nm4000 nm3900065520FIELD EMISSION GUNKodak SO163 film mmObjective lens astigmatism was corrected at 200,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
CartridgeGATAN HELIUM°° eV77 K K K mrad20 e/Å217-OCT-2006
Processing
Protocol:Multi-particle refinement
Software:SPIDER
CTF correction:By defocus groups
Number of particles:81946
Imposed symmetry:C1
Resolution by author:9 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Several rounds of multireference 3D projection refinement.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
874.7 μm/pixelPRIMESCAN
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails