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Title:Structure of the Escherichia coli ribosomal termination complex with release factor 2.
Authors:Klaholz BP, Pape T, Zavialov AV, Myasnikov AG, Orlova EV, Vestergaard B, Ehrenberg M, van Heel M
Sample:release factor RF2 bound to E. coli ribosomes
Aggregation state:Single particle (14 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: release factor RF2 bound to E. coli ribosomes
Oligomeric state: monomer
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryoteribosome2.3Proteins L7/L12 of the LSU 50S subunit are not seen in the map
2ligandrelease factor 20.040monomertrueIPR004374
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.15 mg/mLpolymix bufferno staining, cryo-EM with holey carbon grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE45%20 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 CM200FEG/ST200 kVFLOOD BEAMBRIGHT FIELD2.1 mm500 nm2900 nm5000048000FIELD EMISSION GUNKodac SO163 film- mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid nitrogen-cooled cryo specimen holder.GATAN LIQUID NITROGENnone°none°nonenone eV100 K100 K102 K mrad10 e/Å2-2001-09-06
Processing
Software:IMAGIC
CTF correction:each particle
Resolution by author:14 Å
Resolution method:FSC: value between 3 sigma and 0.5 cut-off
Processing details:exact filtered back-projection
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
783 μm/pixel12linkPATCHWORK DENSITOMETER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1GIX 1GIY rigid bodybest visual fit using the program OeyeREALrigid body of three individual domains keeping the connectivity; linkers betweenn domains defined based on temperature factors of crystal structure, conserved Gly and Pro residues, proteolytic sites, and global domain architecture
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