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Title:Structure of the E. coli signal recognition particle bound to a translating ribosome.
Authors:C.Schaffitzel, M.Oswald, I.Berger, T.Ishikawa, J.P.Abrahams, H.K.Koerten, R.I.Koning, N.Ban
Sample:E. coli ribosome nascent chain complexes and signal recognition particle
Aggregation state:Single particle (15.5 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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Sample
Sample name: E. coli ribosome nascent chain complexes and signal recognition particle
Oligomeric state: One SRP binds to one ribosome.
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
2nucleic-acidA-site tRNAfalse
3nucleic-acidP-site tRNAfalse
4nucleic-acidE-site tRNAfalse
5ligandE. coli SRPmonomertrue
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.25 mg/mL50 mM Hepes-KOH pH 7.5, 100 mM KCl, 25 mM MgCl2, 1 mM DTT, 1 mM GTPcarbon-coated lacey formvar grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%298 KHOMEMADE PLUNGERblot for 1.5 sec before plunging msVitrification instrument: home-built environmental chamber and vitrification device
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2 mm1000 nm4500 nm5000051000FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
cryo stageGATAN LIQUID NITROGEN°° eV K K K mrad10 e/Å2
Processing
Software:Spider
Resolution by author:15.5 Å
Resolution method:FSC at 0.5 cut-off
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
251 μm/pixellinkOTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2IY3 normal mode based flexible fittingcorrelation coefficientNOMADREALFitting of SRP atomic model into isolated EM density
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