top_cornerHomeEMD-1095  Contact us 
Image unavailable
Title:A mutant chaperonin with rearranged inter-ring electrostatic contacts and temperature-sensitive dissociation.
Authors:B. T. Sewell, R. B. Best, S. Chen, A. M. Roseman, G.W. Farr, A.R. Horwich, H.R. Saibil
Sample:E461K mutant of GroEL
Aggregation state:Single particle (25 angstroms resolution)
Red flagLatest update:2011-05-26
inner_corner
tab_top_blue.gif
bullet_white.gif  Summary
tab_bottom_blue.gif
pixel.gif
tab_top_white.gif
bullet.png Experimental details
tab_bottom_white.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Visualization
tab_bottom_blue.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Map information
tab_bottom_blue.gif
pixel.gif
tab_top_blue.gif
bullet_white.gif  Downloads
tab_bottom_blue.gif
pixel.gif
Sample
Sample name: E461K mutant of GroEL
Oligomeric state: homo tetradecamer
Theoretical molecular weight of the sample: 0.84
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinE461K mutant of GroEL0.84homo tetradecamertrueEscherichia colihomo tetradecamer
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
7.40.84 mg/mL20 mM Tris, 5 mM KCl, 10 mM MgCl2, pH 7.4grids were flash frozen by plunging into liquid ethane at -170 degrees C. No stain was used.copper grid, holey carbon films
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGERgrids were blotted prior to plunging for approximately two seconds msVitrification instrument: Home made plunger
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2010HT200 kVFLOOD BEAMBRIGHT FIELD mm1000 nm2000 nm3000030000TUNGSTEN HAIRPINKodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Oxford CT3500OTHER°° eV100 K100 K100 K mrad10 e/Å2
Processing
Software:Spider
Resolution by author:25 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Seven fold symmetry was imposed on the reconstruction.
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
710 μm/pixel16linkScanned on a Leafscan 45 at 2540 pixels per inch. Subsequently pixels were averaged in 2x2 blocks.OTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1EMS The seven membered rings of GroEL (1EMS) were docked separately into the reconstructed density. This was done manually using the programme O.
outer_corner