Benomyl and Colchicine Synergistically Inhibit Cell Proliferation and Mitosis: Evidence of Distinct Binding Sites for These Agents in Tubulin
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Title |
Benomyl and Colchicine Synergistically Inhibit Cell Proliferation and Mitosis: Evidence of Distinct Binding Sites for These Agents in Tubulin
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Creator |
CLEMENT, MJ
RATHINASAMY, K ADJADJ, E TOMA, F CURMI, PA PANDA, D |
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Subject |
anthelmintic benzimidazoles
microtubule dynamics nmr-spectroscopy structural basis brain tubulin bovine brain drugs carbendazim cancer polymerization |
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Description |
Benomyl, a tubulin-targeted antimitotic antifungal agent, belongs to the benzimidazole group of compounds, which are known to inhibit the binding of colchicine to tubulin. Therefore, benomyl was thought to bind at or near the colchicine-binding site on tubulin. However, recent mutational studies in yeast and fluorescence studies involving competitive binding of benomyl and colchicine on goat brain tubulin suggested that benomyl may bind to tubulin at a site distinct from the colchicine-binding site. We set out to examine whether colchicine and benomyl bind to tubulin at distinct sites using a human cervical cancer (HeLa) cell line with the thinking that these agents should exert either additive or synergistic activity on cell proliferation if their binding sites on tubulin are different. We found that benomyl and colchicine synergistically inhibited the proliferation of HeLa cells and blocked their cell cycle progression at mitosis. The synergistic activity of benomyl and colchicine was also apparent from their strong depolymerizing effects on both the spindle and interphase microtubules when used in combinations, providing further evidence that these agents bind to tubulin at different sites. Using NMR spectroscopy, we finally demonstrated that benomyl and colchicine bind to tubulin at different sites and that the binding of colchicine seems to positively influence the binding of benomyl to tubulin and vice versa. Further, an analysis of the saturation transfer difference NMR data yielded an interesting insight into the colchicine-tubulin interaction. The data presented in this study provided a mechanistic understanding of the synergistic effects of benomyl and colchicine on HeLa cell proliferation.
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Publisher |
AMER CHEMICAL SOC
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Date |
2011-07-13T17:46:51Z
2011-12-26T12:48:00Z 2011-12-27T05:43:04Z 2011-07-13T17:46:51Z 2011-12-26T12:48:00Z 2011-12-27T05:43:04Z 2008 |
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Type |
Article
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Identifier |
BIOCHEMISTRY, 47(49), 13016-13025
0006-2960 http://dx.doi.org/10.1021/bi801136q http://dspace.library.iitb.ac.in/xmlui/handle/10054/3732 http://hdl.handle.net/10054/3732 |
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Language |
en
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