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Title:Three-dimensional structures of translating ribosomes by Cryo-EM.
Authors:Gilbert RJ, Fucini P, Connell S, Fuller SD, Nierhaus KH, Robinson CV, Dobson CM, Stuart DI
Sample:70S E. coli ribosome
Aggregation state:Single particle (13.4 angstroms resolution)
Red flagLatest update:2011-05-26
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bullet_white.gif  Summary
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bullet.png Experimental details
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bullet_white.gif  Visualization
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bullet_white.gif  Map information
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bullet_white.gif  Downloads
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Sample
Sample name: 70S E. coli ribosome
Oligomeric state: 30S subunit and 50S subunit
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryoteE. coli 30S subunit
2ribosome-prokaryoteE. coli 50S subunit
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
5.0 mg/mL20 mM HEPES, 150 mM ammonium acetate, 6 mM magnesium acetate, 2 mM spermidine, 0.05 mM spermine and 4 mM 2-mercaptoethanol. Concentration of ribosomes expressed to A260 units.300 mesh copper grid with holey carbon film
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%100 KStandard unmodified guillotine plunger ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG200 kVOTHEROTHER2 mm1475 nm3915 nm50000FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
EucentricGATAN LIQUID NITROGEN°° eV100 K K K mrad e/Å22000-05-01
Processing
Software:SPIDER, IMAGIC, GAP, CNS, XPLOR
CTF correction:Each negative dataset
Resolution by author:13.4 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Final maps were calculated from 12 individual datasets, with scaling in reciprocal space to crystallographic ribosome structures and correction for map anisotropy by B-factor weighting of amplitudes in XPLOR.
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
127 μm/pixel58linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
Rigid bodyR-factor and correlation coefficientGAPREAL
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