Quantification of Engagement of Microtubules by Small Molecules in Living Cells by Flow Cytometry.

ABSTRACT: Drugs such as paclitaxel (Taxol) that bind microtubules are widely used for the treatment of cancer. Measurements of the affinity and selectivity of these compounds for their targets are largely based on studies of purified proteins, and only a few quantitative methods for the analysis of interactions of small molecules with microtubules in living cells have been reported. We describe here a novel method for rapidly quantifying the affinities of compounds that bind polymerized tubulin in living HeLa cells. This method uses the fluorescent molecular probe Pacific Blue-GABA-Taxol in conjunction with verapamil to block cellular efflux. Under physiologically relevant conditions of 37 °C, this combination allowed quantification of equilibrium saturation binding of this probe to cellular microtu

SUBMITTER: Andres AE 

PROVIDER: S-EPMC9585582 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature