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    Structure of microtubule-trapped Human Kinesin-5 and its mechanism of inhibition revealed using Cryoelectron Microscopy

    Peña, A. and Sweeney, A. and Cook, A.D. and Topf, Maya and Moores, Carolyn A. (2020) Structure of microtubule-trapped Human Kinesin-5 and its mechanism of inhibition revealed using Cryoelectron Microscopy. Structure , ISSN 0969-2126. (In Press)

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    Kinesin-5 motors are vital mitotic spindle components, and disruption of their function perturbs cell division. We investigated the molecular mechanism of the human kinesin-5 inhibitor GSK-1, which allosterically promotes tight microtubule binding. GSK-1 inhibits monomeric human kinesin-5 ATPase and microtubule gliding activities, and promotes the motor's microtubule stabilization activity. Using cryoelectron microscopy, we determined the 3D structure of the microtubule-bound motor-GSK-1 at 3.8 Å overall resolution. The structure reveals that GSK-1 stabilizes the microtubule binding surface of the motor in an ATP-like conformation, while destabilizing regions of the motor around the empty nucleotide binding pocket. Density corresponding to GSK-1 is located between helix-α4 and helix-α6 in the motor domain at its interface with the microtubule. Using a combination of difference mapping and protein-ligand docking, we characterized the kinesin-5-GSK-1 interaction and further validated this binding site using mutagenesis. This work opens up new avenues of investigation of kinesin inhibition and spindle perturbation.


    Item Type: Article
    Keyword(s) / Subject(s): kinesin, microtubule, cryo-electron microscopy, image reconstruction, inhibitor, antimitotic
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Depositing User: Administrator
    Date Deposited: 17 Jan 2020 07:48
    Last Modified: 02 Aug 2020 02:35


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