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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 name: Antiporter SOS1
Oligomeric state: Dimer
Theoretical molecular weight of the sample: 0.254
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
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
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
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
Protocol:3D maximum likelihood
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
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails