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Title:Microscopic evidence for a minus-end-directed power stroke in the kinesin motor ncd.
Authors:Wendt TG
Sample:SH3 tagged Ncd296 motor fragment from Drosophila Melanogaster no nucleotide state
Aggregation state:Helical
Red flagLatest update:2012-03-07
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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: SH3 tagged Ncd296 motor fragment from Drosophila Melanogaster no nucleotide state
Oligomeric state: SH3ncd forms a dimer
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinSH3 tagged Ncd296 dimer0.1068SH3ncd296 forms a dimertrue
2proteintubulin dimer0.1polymer of alpha beta dimersfalsebovine tubulin was purchased from Cytoskeleton Inc. (Denver,CO)
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
6.82.45 mg/mL80mM PIPES, 2mM MgCl2plunge-frozen in liquid ethanequantifoil holey grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE%93 KHOMEMADE PLUNGER msVitrification instrument: standard guillotine. vitrification carried out at room temperature
Imaging:
MicroscopeVoltageIllumination modeImaging modeCs:Defocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI/PHILIPS CM200FEG/ST160 kVFLOOD BEAMBRIGHT FIELD1.26 mm1500 nm2000 nm38000TUNGSTEN HAIRPINKodak SO163 film mmobjective lens astigmatism was corrected at 150K mag

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
side entryGATAN LIQUID NITROGEN0°0° eV93 K K K mrad e/Å2low dose imaging
Processing
Software:PHOELIX
Processing details:final map was calculated from 27 datasets of near and far sides
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
1421 μm/pixel0.98linkZEISS SCAI
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
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