+Search query
-Structure paper
Title | Structure of Microtubule-Trapped Human Kinesin-5 and Its Mechanism of Inhibition Revealed Using Cryoelectron Microscopy. |
---|---|
Journal, issue, pages | Structure, Vol. 28, Issue 4, Page 450-457.e5, Year 2020 |
Publish date | Apr 7, 2020 |
Authors | Alejandro Peña / Aaron Sweeney / Alexander D Cook / Julia Locke / Maya Topf / Carolyn A Moores / |
PubMed Abstract | 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 ...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. |
External links | Structure / PubMed:32084356 / PubMed Central |
Methods | EM (helical sym.) |
Resolution | 3.8 - 6.1 Å |
Structure data | EMDB-10421: Human kinesin-5 motor domain in the GSK-1 state bound to microtubules EMDB-10422, PDB-6ta4: |
Chemicals | ChemComp-G2P: ChemComp-MG: ChemComp-MZK: ChemComp-GTP: ChemComp-ANP: |
Source |
|
Keywords | CELL CYCLE / kinesin / microtubule / mitosis / inhibition / motor |