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Title:Incorporation of aminoacyl-tRNA into the ribosome as seen by cryo-electron microscopy.
Authors:Valle M, Zavialov A, Li W, Stagg SM, Sengupta J, Nielsen RC, Nissen P, Harvey SC, Ehrenberg M, Frank J
Sample:70S ribosome from E. coli complex. 70S-tRNAfMet-MF-tRNAPhe
Aggregation state:Single particle (9 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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Sample
Sample name: 70S ribosome from E. coli complex. 70S-tRNAfMet-MF-tRNAPhe
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote-70S from E. coli
2nucleic-acid-fMet-tRNAfMetfalse
3nucleic-acid-Phe-tRNAPhefalse
4protein-Elongation Factor Tufalsen/a
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.5 mg/mL5 mM potassium phosphate, 5 mM magnesium acetate, 5 mM ammonium chloride, 95 mM potassium chloride, 0.5 mM calcium chloride, 8 mM putrescine, 1 mM spermidine, and 1 mM dithioerythritolcryo-electron microscopy. No stain300 mesh Quantifoil R2/4
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%93 Ktwo-face blotting for 1 second ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm2000 nm4000 nm5000049650FIELD EMISSION GUNKodak SO163 film mmcorrected at 175,000

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
single tilt cryoholderGATAN LIQUID NITROGEN0°0° eV93 K K K mrad15 e/Å2
Processing
Software:Spider
CTF correction:segregation in defocus groups and correction in volumes
Resolution by author:9 Å
Resolution method:FSC cut-off at 0.15
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
5014 μm/pixel1.016linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
rigid bodycorrelation coeficientREALfitting carried out manually in "O"
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