Perturbation of microtubule polymerization by quercetin through tubulin binding: a novel mechanism of its antiproliferative activity.
The dietary flavonoid quercetin has a broad range of biological activities, including potent antitumor activity against several types of tumors. Recently, it has been shown that quercetin inhibits cancer cells proliferation by depleting cellular microtubules and perturbing cellular microtubule functions. However, the direct interactions of quercetin with tubulin and microtubules have not been examined so far. Here, we found that quercetin inhibited polymerization of microtubules and depolymerized microtubules made from purified tubulin in vitro. The binding of quercetin with tubulin was studied using quercetin fluorescence and intrinsic tryptophan fluorescence of tubulin. Quercetin bound to tubulin at a single site with a dissociation constant of 5-7 microM, and it specifically inhibited colchicine binding to tubulin but did not bind at the vinblastine site. In addition, quercetin perturbed the secondary structure of tubulin, and the binding of quercetin stimulated the intrinsic GTPase activity of soluble tubulin. Further, quercetin stabilized tubulin against decay and protected two cysteine residues of tubulin toward chemical modification by 5,5'-dithiobis-2-nitrobenzoic acid. Our data demonstrated that the binding of quercetin to tubulin induces conformational changes in tubulin and a mechanism through which quercetin could perturb microtubule polymerization dynamics has been proposed. The data suggest that quercetin inhibits cancer cells proliferation at least in part by perturbing microtubule functions through tubulin binding.
Gupta
Kamlesh
K
BJM School of Biosciences and Bioengineering, Indian Institute of Technology, Bombay, Powai, Mumbai 400 076, India.
Panda
Dulal
D
D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D002942:Circular Dichroism; D003078:Colchicine / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004228:Dithionitrobenzoic Acid / Q000737:chemistry; D004789:Enzyme Activation / Q000187:drug effects; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006041:Goats; D006131:Growth Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D008870:Microtubules / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D011108:Polymers / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011794:Quercetin / Q000378:metabolism / Q000494:pharmacology*; D012995:Solubility; D013050:Spectrometry, Fluorescence; D013439:Sulfhydryl Reagents / Q000737:chemistry; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*; D050257:Tubulin Modulators; D014747:Vinblastine / Q000378:metabolism
D000817:Anilino Naphthalenesulfonates; D006131:Growth Inhibitors; D011108:Polymers; D013439:Sulfhydryl Reagents; D014404:Tubulin; D050257:Tubulin Modulators; D014747:Vinblastine; C035763:5,5'-bis(8-(phenylamino)-1-naphthalenesulfonate); D004228:Dithionitrobenzoic Acid; D011794:Quercetin; D020558:GTP Phosphohydrolases; D003545:Cysteine; D003078:Colchicine