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Title:3D reconstruction a microtubule decorated with monomeric human kinesin (K349 construct) in complex with ADP.
Authors:C.V.Sindelar,K.H.Downing
Sample:Microtubule decorated with monomeric human kinesin (K349 construct) in complex with ADP.
Aggregation state:Helical (9.1 angstroms resolution)
Red flagLatest update:2011-09-23
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Sample
Sample name: Microtubule decorated with monomeric human kinesin (K349 construct) in complex with ADP.
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism GO identifier InterPro identifier Virus identifier Details
1proteinK349trueunidentified
2proteinMicrotubuletrueunidentified
Experiment
Sample preparation:
pHSample conc.DetailsStainingSample support details
6.8 mg/mL25mM Pipes, 25mM NaCl, 2mM MgCl2, 1mM EGTA300 mesh copper grid with homemade holey carbon
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE% KHOMEMADE PLUNGER msVitrification instrument: Homemade
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
JEOL 4000EX400 kVFLOOD BEAMBRIGHT FIELD4.1 mm700 nm1600 nm60000LAB6Kodak SO163 film mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
EucentricGATAN LIQUID NITROGEN°° eV105 K K K mrad15 e/Å208-JAN-2008
Processing
Software:SPIDER,FREALIGN
CTF correction:done within FREALIGN
Resolution by author:9.1 Å
Resolution method:FSC at 0.5 cut-off
Processing details:Approximately 170,000 asymmetric units were averaged in the final reconstruction.
Unit cell:
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
776.35 μm/pixel16linkNIKON SUPER COOLSCAN 9000
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1bg2 Rigid BodyCross correlationUCSF ChimeraREALRegions in the crystal structure whose structure were found to differ from the EM map were excluded from the fitting procedure. These regions corresponded to the switch loops, helices alpha 3 and alpha 4 (switch II helix), and the neck linker.
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