quick and simple detection technique to assess the binding of antimicrotubule agents to the colchicine-binding site快速而简单的检测技术来评估antimicrotubule特工的绑定colchicine-binding站点.pdfVIP

Biol Proced Online (2010) 12:113– 117 DOI 10.1007/s12575-010-9029-5 Quick and Simple Detection Technique to Assess the Binding of Antimicrotubule Agents to the Colchicine-Binding Site Sébastien Fortin Jacques Lacroix Marie-France Côté Emmanuel Moreau Éric Petitclerc René C.-Gaudreault Received: 29 January 2010 /Accepted: 8 March 2010 /Published online: 8 April 2010 # The Author(s) 2010. This article is published with open access at S Abstract Development of antimitotic binding to the tubulin adduct, resulting in an assay that allows the screening colchicine-binding site for the treatment of cancer is rapidly of new molecules targeting this binding site. expanding. Numerous antimicrotubule agents are prepared every year, and the determination of their binding affinity to Keywords colchicine-binding site inhibitor . taxol-binding . . . tubulin requires the use of purified tubulins and radiolabeled site inhibitor N,N-ethylene-bis(iodoacetamide) EBI ligands. Such a procedure is costly and time-consuming and tubulin affinity assay . antimicrotubule agent therefore is limited to the most promising candidates. Here, we report a quick and inexpensive method that requires only usual laboratory resources to assess the binding of antimicro- 1 Introduction tubules to colchicine-binding site. The method is based on the ability of N,N-ethylene-bis(iodoacetamide) (EBI) to cross- Antimicrotubule agents are important in the management of link in living cells the cysteine residues at position 239 and several cancers treatments notably breast, ovarian, and 354 of β-tubulin, residues which are involved in the lung. Unfortunate