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Title:Visualization of release factor 3 on the ribosome during termination of protein synthesis.
Authors:Klaholz BP, Myasnikov AG, Van Heel M
Sample:70S - RF3 complex E.coli
Aggregation state:Single particle (17.5 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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Sample
Sample name: 70S - RF3 complex E.coli
Oligomeric state: monomer
Theoretical molecular weight of the sample: 2.5
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote70S
2nucleic-acidmessenger RNAtrueEscherichia coli
3nucleic-acidisoleucin transfer RNAfalseEscherichia coli
4proteinrelease factor 3monomertrueEscherichia coliIPR004548PrfC gene
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.15 mg/mLpolymix bufferno staining, cryo-EM with holey carbon grids300 mesh Cu/Rh
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% Khome-made cryo-plungerBlot for 2 seconds before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG200 kVFLOOD BEAMBRIGHT FIELD2.1 mm1000 nm2800 nm5000048000FIELD EMISSION GUNKodak SO163 film mmlens astigmatism was corrected at 60,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid nitrogen-cooled cryo specimen holderGATAN LIQUID NITROGEN°° eV K K K mrad10 e/Å22001-19-12
Processing
Software:IMAGIC
CTF correction:individual particles
Resolution by author:17.5 Å
Resolution method:FSC at 0.14 cut-off
Processing details:exact filtered back-projection
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
443 μm/pixel12linkIMAGE SCIENCE PATCHWORK DENSITOMETEROTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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