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Title:Visualization of ribosome-recycling factor on the Escherichia coli 70S ribosome: functional implications.
Authors:Agrawal, Rajendra K.
Sample:70S-RRF complex of e.coli
Aggregation state:Single particle (12 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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Sample
Sample name: 70S-RRF complex of e.coli
Theoretical molecular weight of the sample: 2.5
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote70S2.48
2proteinRRF0.02monomerfalseEscherichia coli
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.5 mg/mL52 mM Tris-HCL (pH7.5), 25 mM KCL, 5.5 mM NH4Cl, 11 mM Mg(OAc)2 0.3 mM Beta-MEquantifoil grids with holy carbon
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE30%93 KHOMEMADE PLUNGERBlot for 2 seconds before plunging msVitrification instrument: two side blotting plunger
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1400 nm4400 nm5000049696FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Cryo transferGATAN LIQUID NITROGEN0°0° eV93 K K K mrad20 e/Å22002-06-01
Processing
Software:SPIDER
CTF correction:defocus groups
Resolution by author:12 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Final map was calculated from 23 CTF corrected defocus groups
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
7614 μm/pixel1.212linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
Rigid Bodycross correlation coefficientmanualthe RRF was fitted as single rigid body
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