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Title:Electron crystallography reveals the structure of metarhodopsin I.
Authors:J. J. Ruprecht, T. Mielke, R. Vogel, C. Villa, G. F. X. Schertler
Sample:Bovine Metarhodopsin I
Aggregation state:2D crystal (5.5 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: Bovine Metarhodopsin I
Oligomeric state: monomer
Theoretical molecular weight of the sample: 0.039
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinRhodopsin0.039MonomerfalseBos taurusGO:0009586IPR000732Illuminated to form photostationary state
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
70.5 mg/mL20mM HEPES pH 7, 100 mM NaCl, 10 mM MgCl2, 3 mM NaN3, 4 mM DTT, 4 mM mercaptoethanol, 2.5 % (v/v) isopropanol300 mesh molybdenum grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE75%77 KGATAN CRYOPLUNGE 3Blot for 20 sec before plungingVitrified after illumination of specimen msVitrification instrument: Gatan cryoplunge. Vitrification carried out in temperature and humidity-controlled chamber
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F30300 kVSPOT SCANBRIGHT FIELD2.3 mm270 nm1410 nm6000059000FIELD EMISSION GUNKodak SO163 film mmcorrected at 230,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Side entry liquid-nitrogen cooled Gatan holderGATAN LIQUID NITROGEN0°60° eV77 K77 K77 K mrad15 e/Å2
Processing
Software:MRC
CTF correction:Each image
Resolution by author:5.5 Å
Resolution method:Point spread function
Crystal growth details:11 day dialysis at 18 deg C
Unit cell:
abGamma
58.8 Å83.7 Å90°
Plane/space group:P 2 21 21
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
877 μm/pixellinkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1GZM Rigid BodyOREAL
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