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

Nature Communications
Grégory MenchonMichel O Steinmetz

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.

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Citations

Nov 16, 2018·Angewandte Chemie·Pol KarierAlois Fürstner
Jan 21, 2021·Bioorganic & Medicinal Chemistry Letters·Ting ZhuSai-Yang Zhang
May 22, 2020·ACS Medicinal Chemistry Letters·Eleonora ColomboDaniele Passarella
Feb 24, 2021·Proceedings of the National Academy of Sciences of the United States of America·Susan MatthewHendrik Luesch
Jun 19, 2021·Biochemical and Biophysical Research Communications·Wenting LiJinliang Yang
Jul 10, 2021·Journal of the American Chemical Society·Qiuling HuangJiang Li
Nov 18, 2021·Chemistry : a European Journal·Paola MarzulloDaniele Passarella
Jul 31, 2020·Mini Reviews in Medicinal Chemistry·Olagoke Zacchaeus OlatundeCanzhong Lu

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Methods Mentioned

BETA
X-ray
fluorescence microscopy

Software Mentioned

SigmaPlot
MolProbity
EQUIGRA5
Phenix
XDS

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