Cryo-EM structure of a microtubule-bound parasite kinesin motor and implications for its mechanism and inhibition

Cook, Alexander D.; Roberts, Anthony J.; Atherton, Joseph; Tewari, Rita; Topf, Maya; Moores, Carolyn A.

Abstract

Plasmodium parasites cause malaria and are responsible annually for hundreds of thousands of deaths. Kinesins are a superfamily of microtubule-dependent ATPases that play important roles in the parasite replicative machinery, which is a potential target for antiparasite drugs. Kinesin-5, a molecular motor that cross-links microtubules, is an established antimitotic target in other disease contexts, but its mechanism in Plasmodium falciparum is unclear. Here, we characterized P. falciparum kinesin-5 (PfK5) using cryo-EM to determine the motor’s nucleotide-dependent microtubule-bound structure and introduced 3D classification of individual motors into our microtubule image processing pipeline to maximize our structural insights. Despite sequence divergence in PfK5, the motor exhibits classical kinesin mechanochemistry, including ATP-induced subdomain rearrangement and cover neck bundle formation, consistent with its plus-ended directed motility. We also observed that an insertion in loop5 of the PfK5 motor domain creates a different environment in the well-characterized human kinesin-5 drug-binding site. Our data reveal the possibility for selective inhibition of PfK5 and can be used to inform future exploration of Plasmodium kinesins as antiparasite targets.

Citation

Cook, A. D., Roberts, A. J., Atherton, J., Tewari, R., Topf, M., & Moores, C. A. (2021). Cryo-EM structure of a microtubule-bound parasite kinesin motor and implications for its mechanism and inhibition. Journal of Biological Chemistry, 297(5), Article 101063. https://doi.org/10.1016/j.jbc.2021.101063

• Journal Article Type: Article
• Acceptance Date: Jul 23, 2021
• Online Publication Date: Oct 13, 2021
• Publication Date: 2021-11
• Deposit Date: Jun 20, 2025
• Publicly Available Date: Jun 23, 2025
• Journal: Journal of Biological Chemistry
• Print ISSN: 0021-9258
• Electronic ISSN: 1083-351X
• Publisher: American Society for Biochemistry and Molecular Biology
• Peer Reviewed: Peer Reviewed
• Volume: 297
• Issue: 5
• Article Number: 101063
• DOI: https://doi.org/10.1016/j.jbc.2021.101063
• Keywords: ATPase, anti-parasitic, cell division, cryo-EM, kinesin, malaria, mechanochemistry, microtubule, motor, Plasmodium falciparum
• Public URL: https://nottingham-repository.worktribe.com/output/6739436
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0021925821008668?via%3Dihub