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Title:Three-dimensional structure of the 70S E. coli ribosome in its native 3D organization in polysomes.
Authors:F.Brandt,A.H.Elcock,S.A.Etchells,J.O.Ortiz,F.U.Hartl,W.Baumeister
Sample:70S ribosome in a native polysome with a neighbor ribosome at the 5' end oriented in a tt-class
Aggregation state:Single particle (30 angstroms resolution)
Red flagLatest update:2010-11-01
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Sample
Sample name: 70S ribosome in a native polysome with a neighbor ribosome at the 5' end oriented in a tt-class
Oligomeric state: ribosomes forming polysomes
Theoretical molecular weight of the sample: 2.7
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote70S ribosome2.7The ribosomes under scrutiny are part of large polysomal arrangements
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.520 mg/mLRoche reconstitution buffer from the RTS kit, ATP,GTP, 8-15 mM Mg2, 100-250 nm K,NH4,DTT, protease inhibitors,NaN3400 mesh gold grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%77 KHOMEMADE PLUNGERBlot for 1 s before plunging msVitrification instrument: plunger. Vitrification carried out in air
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG160 kVFLOOD BEAMBRIGHT FIELD2 mm3000 nm4000 nm2750053960FIELD EMISSION GUNTietz CCD camera 4Kx4K mmObjective astigmatism was corrected using a quadrupole stigmator at 50,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid nitrogen-GATAN LIQUID NITROGEN-60°60° eV77 K70 K90 K-4 mrad50 e/Å201-Dec-2006
Processing
Software:TOM ToolBox
Resolution by author:30 Å
Resolution method:FSC at 0.5 cut-off
Processing details:CET
Unit cell:
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2AW7 Rigid BodyChimeraREAL
2AWB Rigid BodyChimeraREAL
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