top_cornerHomeEMD-2538  Contact EMDataBank 
Image unavailable
Title:cryo-electron microscopy of microtubule-bound human kinesin-5 motor domain in the ADP state with a gold cluster attached to the loop5 (T126C)
Authors:GOULET A, MAJOR J, JUN Y, GROSS SP, ROSENFELD SR, MOORES CA
Sample:13-protofilament microtubule-bound human kinesin-5 motor domain in presence of ADP
Method:Single particle reconstruction (18 angstroms resolution)
Red flagLatest update:2014-02-12
Sample
Sample name: 13-protofilament microtubule-bound human kinesin-5 motor domain in presence of ADP
Oligomeric state: 13-protofilament microtubule with one kinesin-5 motor domain bound every tubulin heterodimers
Components:
ID Type Name Exp. MW (MDa) Theo. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinalpha tubulinfalseBos taurusQ2HJ86IPR002452
2proteinbeta tubulinfalseBos taurusQ6B856IPR013838
3proteinKinesin-5 motor domainmonomertrueHomo sapiensP52732IPR019821cys-lite mutant containing the substitution T126C. A undecagold cluster is covalently attached to T126C.
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
6.8 mg/mL20 mM PIPES, 5 mM MgCl2, 1 mM EGTA, 10 mM ADP, 2% glycerol400 mesh holey carbon grids
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE100% KFEI VITROBOT MARK Ichamber at 24 degrees C, blot 3.5 sec ms
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI TECNAI F20200 kVFLOOD BEAMBRIGHT FIELD2.0 mm1100 nm2600 nm68000FIELD EMISSION GUNGATAN ULTRASCAN 4000 (4k x 4k) mmObjective lens astigmatism was corrected at 150,000 times magnification

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
GATAN LIQUID NITROGEN°° eV90 K K K mrad18 e/Å216-JAN-2013
Processing
Software:SPIDER, FREALIGN
CTF correction:FREALIGN
Number of particles:1000
Imposed symmetry:C1
Resolution by author:18 Å
Resolution method:FSC 0.5, semi-independent
Processing details:Approximately 135,000 asymmetric units were averaged in the final reconstruction. The particles were selected along individual microtubules.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
20 μm/pixel
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails