Insights into the Interaction of Discodermolide and Docetaxel with Tubulin. Mapping the Binding Sites of Microtubule-Stabilizing Agents by Using an Integrated NMR and Computational Approach

The binding interactions of two antitumor agents that target the paclitaxel site, docetaxel and discodermolide, to unassembled α/β-tubulin heterodimers and microtubules have been studied using biochemical and NMR techniques. The use of discodermolide as a water-soluble paclitaxel biomimetic and exte...

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Bibliographic Details
Published in:ACS chemical biology 2011-08, Vol.6 (8), p.789-799
Main Authors: Canales, Angeles, Rodríguez-Salarichs, Javier, Trigili, Chiara, Nieto, Lidia, Coderch, Claire, Andreu, José Manuel, Paterson, Ian, Jiménez-Barbero, Jesús, Díaz, J. Fernando
Format: Article
Language:English
Subjects:
Alkanes - chemistry
Alkanes - pharmacology
Antineoplastic Agents - chemistry
Antineoplastic Agents - pharmacology
Binding Sites
Carbamates - chemistry
Carbamates - pharmacology
Computer Simulation
Humans
Lactones - chemistry
Lactones - pharmacology
Microtubules - chemistry
Microtubules - metabolism
Models, Molecular
Molecular Conformation
Nuclear Magnetic Resonance, Biomolecular - methods
Paclitaxel - chemistry
Paclitaxel - pharmacology
Protein Multimerization
Pyrones - chemistry
Pyrones - pharmacology
Taxoids - chemistry
Taxoids - pharmacology
Tubulin - chemistry
Tubulin - metabolism
Tubulin Modulators - chemistry
Tubulin Modulators - pharmacology
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Summary:The binding interactions of two antitumor agents that target the paclitaxel site, docetaxel and discodermolide, to unassembled α/β-tubulin heterodimers and microtubules have been studied using biochemical and NMR techniques. The use of discodermolide as a water-soluble paclitaxel biomimetic and extensive NMR experiments allowed the detection of binding of microtubule-stabilizing agents to unassembled tubulin α/β-heterodimers. The bioactive 3D structures of docetaxel and discodermolide bound to α/β-heterodimers were elucidated and compared to those bound to microtubules, where subtle changes in the conformations of docetaxel in its different bound states were evident. Moreover, the combination of experimental TR-NOE and STD NMR data with CORCEMA-ST calculations indicate that docetaxel and discodermolide target an additional binding site at the pore of the microtubules, which is different from the internal binding site at the lumen previously determined by electron crystallography. Binding to this pore site can then be considered as the first ligand-protein recognition event that takes place in advance of the drug internalization process and interaction with the lumen of the microtubules.
ISSN:1554-8929
1554-8937
DOI:10.1021/cb200099u