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Title:Structure of the E. coli protein-conducting channel bound to a translating ribosome.
Authors:K.Mitra,C.Schaffitzel,T.Shaikh,F.Tama,S.Jenni,C.L.Brooks III,N.Ban, J.Frank
Sample:co-translational E. coli 70S ribosome-nascent chain complexed with SecYEG
Aggregation state:Single particle (14.9 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: co-translational E. coli 70S ribosome-nascent chain complexed with SecYEG
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote30S
2ribosome-prokaryote50S
3nucleic-acidA-site tRNAfalse
4nucleic-acidP-site tRNAfalse
5nucleic-acidE-site tRNAfalse
6nucleic-acidmRNAtrue
7proteinprotein-conducting channeldimer of heterotrimertrueunidentified
8proteinnascent chaintrueunidentifiedStrep-II-FtsQ-SecM construct
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
mg/mL
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE90%93 KHOMEMADE PLUNGERBlot for 2 seconds before plunging msVitrification instrument: two sided blotting plunger
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F30300 kVFLOOD BEAMBRIGHT FIELD2.26 mm1500 nm4300 nm3900039000FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Cryo stageOTHER°° eV93 K K K mrad11 e/Å22004-03-09
Processing
Software:SPIDER package
CTF correction:defocus groups
Resolution by author:14.9 Å
Resolution method:FSC at 0.5 cut-off
Processing details:The falloff of Fourier amplitudes toward higher spatial frequencies was corrected using the x-ray solution scattering intensity distribution of 70S ribosomes from E. coli during each round of refinement
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
38514 μm/pixel1.212linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2AKH normal mode-based flexible fittingcorrelation coefficientREALFitting of SecYEG atomic model into isolated EM density of protein-conducting channels
2AKI normal mode-based flexible fittingcorrelation coefficientREALFitting of SecYEG atomic model into isolated EM density of protein-conducting channels
real space refinementR-factorRSR2000REAL
real space refinementR-factorRSR2000REAL
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