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Title:ribosome-RelA complex
Authors:Agirrezabala X, Fernandez I, Kelley A, Gil-Carton D, Ramakrishnan V, Valle M
Sample:70S-RelA complex
Method:Single particle reconstruction (14.5 angstroms resolution)
Red flagLatest update:2013-09-11
Sample
Sample name: 70S-RelA complex
Theoretical molecular weight of the sample: 2.5
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1ribosome-prokaryote70S ribosome2.5falseThermus thermophilus
2nucleic-aciddeacylated tRNAfalseThermus thermophilus
3nucleic-acidtRNAfalseThermus thermophilus
4nucleic-acidtRNAfalseThermus thermophilus
5proteinRelAfalseEscherichia coli
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.450.25 mg/mL5mM Hepes-KOH pH 7.45, 50mM KCl, 10mM NH4Cl, 10mM Mg-acetate, 5mM beta-mercapto-ethanolcryoQuantifoil grids (2/4) with thin carbon on top
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100%90 KFEI VITROBOT MARK IBlot for 3 seconds before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2200FS200 kVFLOOD BEAMBRIGHT FIELD2 mm1000 nm4000 nm4000040000FIELD EMISSION GUNKODAK SO-163 FILM mmcorrected at 100k

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°°in-column Omega filter15-20 eV K K K mrad10 e/Å215-JAN-2012
Processing
Protocol:projection matching
Software:XMIPP, SPIDER
CTF correction:defocus groups
Number of particles:28791
Imposed symmetry:C1
Resolution by author:14.5 Å
Resolution method:FSC 0.5
Processing details:Particles were selected by combining automated particle picking , multivariate data analysis and visual inspection.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
4257 μm/pixel8OTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
2WRN flexibleMDFFREAL
2WRO flexibleMDFFREAL