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Title:Reconfiguration of yeast 40S ribosomal subunit domains by the translation initiation multifactor complex.
Authors:R.J.C. Gilbert, Y. Gordiyenko, T. von der Haar,
Sample:Eukaryotic translation preinitiation complex 43S
Aggregation state:Single particle (30.0 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: Eukaryotic translation preinitiation complex 43S
Oligomeric state: 40S-Met-tRNA-eIF2-GMP-PNP-eIF3-eIF1-eIF1A
Theoretical molecular weight of the sample: 1.941
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-eukaryoteSmall subunit1.4S. cerevisiae
2nucleic-acidtRNA0.023falseS. cerevisiae
3proteineukaryotic translation initiation factor 20.124heterotrimertrueS. cerevisiae
4proteineukaryotic translation initiation factor 30.362HeteropentamertrueS. cerevisiae
5proteineukaryotic translation initiation factor 10.012MonomertrueS. cerevisiae
6proteineukaryotic translation initiation factor 1A0.017MonomertrueS. cerevisiae
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.4 mg/mL38mM HEPES, 135mM KAc, 3.25mM MgAc2, 5mM beta-mercaptoethanol, 10uM GMP-PNP300 mesh copper grid with lacey carbon film
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERBlot with Whatman number 1 paper for 1-2 seconds prior to plunging. msVitrification instrument: Home-made plunger
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG200 kVFLOOD BEAMBRIGHT FIELD2 mm2055 nm11685 nm50000FIELD EMISSION GUNKodak SO163 film mmAstigmatism corrected at 100,000 x

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
EucentricGATAN LIQUID NITROGEN°° eV100 K K K mrad e/Å2
Processing
Software:IMAGIC, EMAN, FREALIGN, SPIDER, GAP
CTF correction:Per micrograph
Resolution by author:30.0 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Final maps were computed from CTF-corrected (by phase flipping) images, and scaled in Fourier space to a scattering model of the structure.
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
708.33 μm/pixel8linkOTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
Rigid bodyReal space CC and R-factorGAPREAL
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