Journal: J Mol Biol / Year: 2009 Title: 9-Angström structure of a microtubule-bound mitotic motor. Authors: Andrew J Bodey / Masahide Kikkawa / Carolyn A Moores / Abstract: Kinesin-5 (K5) motors are important components of the microtubule (MT)-based cell division machinery and are targets for small-molecule inhibitors currently in cancer clinical trials. However, the ...Kinesin-5 (K5) motors are important components of the microtubule (MT)-based cell division machinery and are targets for small-molecule inhibitors currently in cancer clinical trials. However, the nature of the K5-MT interaction and the regulatory mechanisms that control it remain unclear. Using cryo-electron microscopy and image processing, we calculated the structure of a K5 motor bound to MTs at 9 A resolution, providing insight into this important interaction. Our reconstruction reveals the K5 motor domain in an ATP-like conformation in which MT binding induces the conserved nucleotide-sensing switch I and II loops to form a compact subdomain around the bound nucleotide. Our reconstruction also reveals a novel conformation for the K5-specific drug-binding loop 5, suggesting a possible role for it in switching K5s between force generation and diffusional modes of MT binding. Our data thus shed light on regulation of the interaction between spindle components important for chromosome segregation.
History
Deposition
Feb 26, 2009
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Header (metadata) release
Mar 9, 2009
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Map release
Apr 1, 2009
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Update
Mar 13, 2013
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Current status
Mar 13, 2013
Processing site: PDBe / Status: Released
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Structure visualization
Movie
Surface view with section colored by density value
Cryogen name: ETHANE / Instrument: HOMEMADE PLUNGER / Details: Vitrification instrument: Home made / Method: Blot for 2-3 seconds before plunging
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Electron microscopy
Microscope
FEI TECNAI F20
Alignment procedure
Legacy - Astigmatism: Astigmatism corrected at 150,000 times magnification
Image recording
Category: FILM / Film or detector model: KODAK SO-163 FILM / Digitization - Scanner: ZEISS SCAI / Digitization - Sampling interval: 7.0 µm / Number real images: 24 / Average electron dose: 10 e/Å2 / Bits/pixel: 12
Electron beam
Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
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