top_cornerHomeEMD-2182  Contact EMDataBank 
Image unavailable
Title:Electron microscopy map of SOS1 antiporter
Authors:Nunez Ramirez R, Sanchez Barrena MJ, Villalta I, Vega JF, Pardo JM, Quintero FJ, Martinez Salazar J, Albert A
Sample:Antiporter SOS1
Method:Single particle reconstruction (25.9 angstroms resolution)
Red flagLatest update:2012-11-28
Sample
Sample name: Antiporter SOS1
Oligomeric state: Dimer
Theoretical molecular weight of the sample: 0.254
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinNa+/H+ antiporter SOS1DimertrueArabidopsis thaliana
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
8.5 mg/mL50 mM phosphate buffer pH 8.5, 300 mM NaCl, 10% glycerol, 0.05% DDM, 250 mM imidazolePurified SOS1 was adsorbed to glow-discharged carbon coated grids and stained with 2% uranyl formate400 mesh carbon coated copper grid, glow discharged in moderate vacuum
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NONE% KNONE ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 2100200 kVFLOOD BEAMBRIGHT FIELD1.0 mm1000 nm2500 nm2000020000LAB6GATAN ORIUS SC200 (2k x 2k) mmObjective lens astigmatism was corrected at 120.000-150.000 times magnification and confirmed by inspection of the Fourier transform

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
JEOL eV K K K mrad15 e/Å201-NOV-2010
Processing
Protocol:3D maximum likelihood
Software:XMIPP
CTF correction:Each micrograph. Estimation using CTFFIND3 and correction using Bsoft
Number of particles:6300
Number of class averages:96
Imposed symmetry:C2
Resolution by author:25.9 Å
Resolution method:FSC 0.5
Processing details:Single images of SOS1 were manually extracted using EMAN. 2D reference-free classification, 2D averaging, 3D classification and reconstruction were performed using maximum-likelihood methods implemented in XMIPP package.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
90 μm/pixel14
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails