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A fluorescence anisotropy assay to discover and characterize ligands targeting the maytansine site of tubulin.

Published version
Peer-reviewed

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Authors

Menchon, Grégory 
Prota, Andrea E 
Lucena-Agell, Daniel 
Bucher, Pascal 
Jansen, Rolf 

Abstract

Microtubule-targeting agents (MTAs) like taxol and vinblastine are among the most successful chemotherapeutic drugs against cancer. Here, we describe a fluorescence anisotropy-based assay that specifically probes for ligands targeting the recently discovered maytansine site of tubulin. Using this assay, we have determined the dissociation constants of known maytansine site ligands, including the pharmacologically active degradation product of the clinical antibody-drug conjugate trastuzumab emtansine. In addition, we discovered that the two natural products spongistatin-1 and disorazole Z with established cellular potency bind to the maytansine site on β-tubulin. The high-resolution crystal structures of spongistatin-1 and disorazole Z in complex with tubulin allowed the definition of an additional sub-site adjacent to the pocket shared by all maytansine-site ligands, which could be exploitable as a distinct, separate target site for small molecules. Our study provides a basis for the discovery and development of next-generation MTAs for the treatment of cancer.

Description

Keywords

Ado-Trastuzumab Emtansine, Animals, Antineoplastic Agents, Binding Sites, Fluorescence Polarization, Humans, Ligands, Macrolides, Maytansine, Microtubules, Oxazoles, Trastuzumab, Tubulin

Journal Title

Nat Commun

Conference Name

Journal ISSN

2041-1723
2041-1723

Volume Title

9

Publisher

Springer Science and Business Media LLC