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Closed Access item Characterizing Ligand-Microtubule binding by competition methods

Authors:Díaz, José Fernando
Martínez Buey, Rubén
Keywords:Microtubules, paclitaxel, epothilones, stopped flow, fluorescence, anisotropy
Issue Date:2007
Publisher:Jun Zhou
Citation:Microtubule Protocols (Methods in Molecular Medicine), 137 (J. Zhou, Ed.) Humana Press, pp 245-260
Abstract:The knowledge of the thermodynamics and kinetics of drug-microtubule interaction is essential to understand the structure/affinity relationship of a given ligand family. When a ligand does not show an appropriate signal change (absorbance or fluorescence) upon binding, the extensive direct characterization of its binding affinities and kinetic rate constants of association and dissociation becomes a complex task. In those cases it is possible to obtain these parameters by competition of the ligand with a reference one of the same binding site that shows such change. Nevertheless, although the experimental setup of the competition measurements is easier, the treatment of the data is complex because simultaneous equilibrium/kinetic equations have to be solved. In this chapter, the taxoid-binding site of the microtubules will be used as an example to describe experimental competition and data analysis methods to determine the binding constants and kinetic rates of association and dissociation of ligands for microtubules
Description:16 páginas, 2 figuras, 1 tabla -- PAGS nros. 245-260
Publisher version (URL):http://dx.doi.org/ 10.1007/978-1-59745-442-1_17
E-ISSNmetadata.dc.identifier.doi = DOI:1940-6037
Appears in Collections:(CIB) Artículos

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