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Title:ATP-bound states of GroEL captured by cryo-electron microscopy.
Authors:Ranson NA, Farr GW, Roseman AM, Gowen B, Fenton WA, Horwich AL, Saibil HR
Sample:GroEL-ATP from E.coli
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: GroEL-ATP from E.coli
Oligomeric state: 14-mer
Theoretical molecular weight of the sample: 0.8
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1proteinGroEL0.814-mertrueE. coliD398A mutant; The Asp398Ala mutant of GroEL has severely reduced ATPase activity but can support a round of protein folding.
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.8 mg/mL12.5 mM HEPES, 5 mM KCl, 5 mM MgCl2, 250 microM ATPholey carbon film
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%100 Kself madeBlot for 1 second before plunging ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG/ST200 kVFLOOD BEAMBRIGHT FIELD2 mm1200 nm5000 nm3800036080FIELD EMISSION GUNKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
EucentricGATAN LIQUID NITROGEN°° eV105 K K K mrad20 e/Å2
Processing
Software:Spider
CTF correction:CTF multiplication and merging of 2D averages
Resolution by author:14.9 Å
Resolution method:FSC at 0.5
Processing details:Filtered back projection
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
1607 μm/pixel18linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1DER Rigid bodyDockEMREALManual fitting of the 3 domains of a subunit as rigid bodies using O, followed by refinement with DockEM
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