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Title:Clathrin D6 coat with auxilin J-domain
Authors:Fotin A, Cheng Y, Sliz P, Grigorieff N, Harrison SC, Kirchhausen T, Walz T
Sample:CLATHRIN COATS BOUND WITH AUXILIN(547-910)
Method:Single particle reconstruction (12 angstroms resolution)
Red flagLatest update:2010-05-05
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Sample
Sample name: CLATHRIN COATS BOUND WITH AUXILIN(547-910)
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinclathrin coat bound with AuxilinD6 assemblefalseBos taurus
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.01 mg/mL20 mM Hepes, pH7.0, 2mM MgCl2,25mM KCl, 10mM (NH4)2SO4holey carbon grid
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%93 KFEI VITROBOT msVitrification instrument: FEI Vitrobot
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2 mm2000 nm5000 nm5000051159FIELD EMISSION GUNKODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
side entry liquid nitrogen-cooled cryo specimen holderGATAN LIQUID NITROGEN°° eV93 K K K mrad e/Å2
Processing
Protocol:common lines
Software:Frealign
CTF correction:CTF correction of each particle.
Number of particles:900
Imposed symmetry:D6
Resolution by author:12 Å
Resolution method:FSC 0.143
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
1907 μm/pixel1.48ZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1N4C 1BPO 1B89 rigid bodyDENSITY CORRELATIONOREALProtocol: Rigid Body. various segment of clathrin heavy chain were separately fitted by manual docking using program O, and fitting was improved by MAVE