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Title:Conformational transition of initiation factor 2 from the GTP- to GDP-bound state visualized on the ribosome.
Authors:A. G. Myasnikov, S.Marzi, A. Simonetti, A.M. Giuliodori, C. O. Gualerzi, G. Yusupova, M. Yusupov, B. P. Klaholz
Sample:Ribosome complex with initiation factor 2 in GTP state
Aggregation state:Single particle (11.5 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: Ribosome complex with initiation factor 2 in GTP state
Oligomeric state: monomer
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-prokaryote70ST.thermophilus; Thermus thermophilus 70S ribosome
2nucleic-acidmessenger RNAtrue
3nucleic-acidformyl-methionyl-RNAfalseTermus Termophilus
4proteininitiation factor 2monomertrueThermus thermophilusGO:0006413IPR000178T.thermophilus; Factor with GTP analogue - GMPPCP
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.5 mg/mL20mM Tris-HCl (pH7.5) ,20mM MgCl2, 100mM KCl, 1mM DTTno staining, cryo-EM with holey carbon grids300 mesh Copper/Rhodium grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERBlot for 2 seconds before plunging msVitrification instrument: home-made cryo-plunger
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1000 nm2400 nm50000FIELD EMISSION GUNKodak SO163 film mmlens astigmatism was corrected at 50,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
ide entry liquid nitrogen-cooled cryo specimen holderGATAN LIQUID NITROGEN°° eV77 K K K mrad10 e/Å22004-03-11
Processing
Software:IMAGIC
CTF correction:individual particles
Resolution by author:11.5 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
363 μm/pixel16linkHeidelberg DruckmaschinenPRIMESCAN
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
Rigid Bodyprogram OREAL
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