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Title:Cryo-EM structure of gastric H+,K+-ATPase with bound Rb+
Authors:Abe K, Tani K, Friedrich T, Fujiyoshi Y
Sample:H+,K+-ATPase bound with Rb+
Method:2D crystallography (8.0 angstroms resolution)
Sample
Sample name: H+,K+-ATPase bound with Rb+
Oligomeric state: One alpha and one beta chain of HK-ATPase
Theoretical molecular weight of the sample: 0.15
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinPOTASSIUM-TRANSPORTING ATPASE0.15DimerfalseSus scrofaGO:0006754IPR008250
Experiment
Specimen state: 2D array
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
4.88 mg/mL20mM propionate, 1mM ADP, 3mM DTT,pH 4.8-4.9 adjusted by Tris, 1mM MgCl2, 1mM AlCl3, 4mM NaF, 10mM RbClmolybdenum grid with thin carbon support
Crystal growth details:Crystals grown in dialysis
Plane group:P 2 21 21
Unit cell:
abGamma
141.0 Å110.6 Å90.0°
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
NITROGEN% KLEICA KF80 msVitrification carried out in cold room at 4 degrees Celsius
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL KYOTO-3000SFF300 kVFLOOD BEAMBRIGHT FIELD1.6 mm830 nm3480 nm4000039500FIELD EMISSION GUNKODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Helium cooledJEOL-60°60° eV K K K mrad e/Å223-MAR-2010
Processing
Software:MRC
CTF correction:Each image
Resolution by author:8.0 Å
Processing details:Images were processed using MRC suite.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
2487 μm/pixel12ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails