REL PDBe PDBe PDBe 2012-04-19 2012-07-09 2012-11-21 2013-01-09 Electron cryo-microscopy of microtubule-bound human kinesin-5 motor domain in rigor state (gold cluster in loop5 T126C). Goulet A Behnke-Parks WM Sindelar CV Major J Rosenfeld SS Moores CA electron cryo-microscopy, kinesin, microtubule, mitosis, cancer Goulet A Behnke-Parks WM Sindelar CV Major J Rosenfeld SS Moores CA The structural basis of force generation by the mitotic motor kinesin-5. J.BIOL.CHEM. 287 44654 44666 2012 23135273 doi:10.1074/jbc.M112.404228 13-protofilament microtubule-bound human kinesin-5 motor domain in absence of nucleotides. A gold cluster is attached to loop5 (T126C). 13-protofilament microtubule-bound human kinesin-5 motor domain in absence of nucleotides. A gold cluster is attached to loop5 (T126C). 13-protofilament microtubule with one kinesin-5 motor domain bound every tubulin heterodimers 3 alpha tubulin Bos taurus Cattle brain heterodimer false IPR002452 beta tubulin Bos taurus brain heterodimer false IPR013838 Kinesin-5 motor domain Homo sapiens Human
undecagold cluster was attached to the specific cysteine residue in loop5 (T126C)
monomer true Escherichia coli pET21a IPR019821 singleParticle particle 6.8
80 mM PIPES, 5 mM MgCl2, 1 mM EGTA, 1 U/mL apyrase
400 mesh holey carbon grids
 ETHANE 100 FEI VITROBOT MARK I chamber at 24 degrees C, blot 2.5 sec FEI TECNAI F20 FLOOD BEAM BRIGHT FIELD FIELD EMISSION GUN 200 2.0 1.2 2.4 50000.0 GATAN LIQUID NITROGEN 90 Objective lens astigmatism was corrected at 150,000 times magnification 2011-11-09 KODAK SO-163 FILM ZEISS SCAI 7 30 18 8.
The particles were selected along individual microtubules.
FREALIGN
 11.0 FSC 0.5 CUT-OFF SPIDER, FREALIGN
Approximately 23,000 asymmetric units were averaged in the final reconstruction. The deposited map is low pass filtered at 16 A.
2027 emd_2079.map.gz 1 IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) 45 45 45 -28 -14 -42 45 45 45 126.0 126.0 126.0 90.0 90.0 90.0 X Y Z -6.72172165 7.19623137 0.23717062 1.92618001 2.8 2.8 2.8 1.37 AUTHOR 3D reconstruction of microtubule-bound human kinesin-5 motor domain with an empty nucleotide-binding site and a gold cluster attached to loop5 (T126C).
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emd_2079.jpg