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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:GroES-ADP7-GroEL-ATP7 from E.coli
Aggregation state:Single particle (23.5 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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bullet_white.gif  Visualization
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Sample
Sample name: GroES-ADP7-GroEL-ATP7 from E.coli
Oligomeric state: 14-mer
Theoretical molecular weight of the sample: 1
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Mutant Organism GO identifier InterPro identifier Virus identifier Details
1proteinGroEL0.814-mertrueE. coli
2proteinGroES0.077-mertrueE. coli
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.50.8 mg/mL12.5 mM HEPES, 5 mM KCl, 5 mM MgCl2, 100 uM ADP, then 2x molar excess of single ring mutant of GroEL (SR1), then 1 mM ATP just before vitrification. See sample details for an explanation of the experimental design.holey carbon film
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%100 Kself madeBlot for 1 second before plungingThe sample was vitrified within a few seconds of manually mixing in ATP. ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2 mm900 nm3000 nm50000FIELD 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:23.5 Å
Resolution method:FSC at 0.5
Processing details:Filtered back projection
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
7114 μm/pixel18linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1AON Rigid bodyManual fitting in OREALThe only movement relative to the crystal structure of GroEL-ES-ADP seen at this resolution is a twist of the free apical domains.
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