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Title:Nucleotide-induced conformations in the neck region of dimeric kinesin.
Authors:Georgios Skiniotis
Sample:Rat kinesin dimer with an SH3 domain complexed to subtilisin treated microtubules in the presence of AMP-PNP.
Aggregation state:Helical (25 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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bullet_white.gif  Map information
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Sample
Sample name: Rat kinesin dimer with an SH3 domain complexed to subtilisin treated microtubules in the presence of AMP-PNP.
Oligomeric state: dimer
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinrat kinesin0.98dimertrueRattus norvegicus
2proteintubulin0.11hetero-dimerfalseRattus norvegicus
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
6.80.5 mg/mLPIPES 80 mM, MgCl2 1 mM, GTP 1 mM, Taxol 20 uM, DMSO 7.5%ice-embededholey grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%93 K ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG/ST200 kVFLOOD BEAMBRIGHT FIELD2 mm2000 nm2500 nm38000FIELD EMISSION GUNKodak SO163 film mmwas corrected at 180,000 times mag.

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
side entryGATAN LIQUID NITROGEN0°0° eV K K K mrad5 e/Å2
Processing
Software:PHOELIX, SUPRIM
Resolution by author:25 Å
Resolution method:FSC at 0.5
Processing details:Final map from 24 averaged datasets = 12 helical tubes
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
1221 μm/pixel8linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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