top_cornerHomeEMD-1006  Contact us 
Image unavailable
Title:A cryo-electron microscopic study of ribosome-bound termination factor RF2.
Authors:U. Rawat, H.Gao, A. Zavialov, R. Gursky, M. Ehrenberg, J. Frank
Sample:E.coli 70s ribosome
Aggregation state:Single particle (11.3 angstroms resolution)
Red flagLatest update:2011-05-26
inner_corner
tab_top_blue.gif
bullet_white.gif  Summary
tab_bottom_blue.gif
pixel.gif
tab_top_white.gif
bullet.png Experimental details
tab_bottom_white.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Visualization
tab_bottom_blue.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Map information
tab_bottom_blue.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Downloads
tab_bottom_blue.gif
pixel.gif
Sample
Sample name: E.coli 70s ribosome
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-prokaryoteE.coli 70s ribosome
2ligandpeptidyl P-tRNAfalse
3ligandE-tRNAfalse
4ligandMFTI-mRNAtrueZavialov et al., A posttermination ribosomal complex is the guanine nucleotide exchange factor for peptide release factor RF3. Cell. 107,1-20 (2001).
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.09 mg/mLPolymix buffer, containing at final concentration 5 mM potassium phosphate, 5 mM magnesium acetate, 5mM ammonium chloride, 95 mM potassium chloride, 0.5 mM calcium chloride, 8mM putrescine, 1mM dithioerythritol
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE58%36 KHOMEMADE PLUNGERBlot and Plunge msVitrification instrument: Two side blotting plunger. Rapid-freezing in liquid ethane
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1390 nm5975 nm5000049696FIELD EMISSION GUNKodac SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Oxford,cryo-transfer 3500GATAN LIQUID NITROGEN0°0° eV93 K K K mrad20 e/Å22001-08-09
Processing
Software:SPIDER/WEB
CTF correction:Wiener filtration of defocus groups
Resolution by author:11.3 Å
Resolution method:FSC at 0.5 cutoff
Processing details:Frank, J. (2000) Three-Dimensional Cryoelectron Microscopy of Ribosomes, Methods of Enzymology (Ch.18) 317, 276-291.
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
3414 μm/pixel12linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1M16 1MVR manual fitting using O
outer_corner