title: 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.
abstract: 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.
authors.affiliation: Dept. Chemical and Physical Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, 28040 Madrid, Spain.
authors.forename: Angeles
authors.initials: A
authors.lastname: Canales
authors.forename: Javier
authors.initials: J
authors.lastname: Rodríguez-Salarichs
authors.forename: Chiara
authors.initials: C
authors.lastname: Trigili
authors.forename: Lidia
authors.initials: L
authors.lastname: Nieto
authors.forename: Claire
authors.initials: C
authors.lastname: Coderch
authors.forename: José Manuel
authors.initials: JM
authors.lastname: Andreu
authors.forename: Ian
authors.initials: I
authors.lastname: Paterson
authors.forename: Jesús
authors.initials: J
authors.lastname: Jiménez-Barbero
authors.forename: J Fernando
authors.initials: JF
authors.lastname: Díaz
mesh_terms: D000473:Alkanes / Q000737:chemistry / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D001665:Binding Sites; D002219:Carbamates / Q000737:chemistry / Q000494:pharmacology*; D003198:Computer Simulation; D000077143:Docetaxel; D006801:Humans; D007783:Lactones / Q000737:chemistry / Q000494:pharmacology*; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008968:Molecular Conformation; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017239:Paclitaxel / Q000737:chemistry / Q000494:pharmacology; D055503:Protein Multimerization; D011753:Pyrones / Q000737:chemistry / Q000494:pharmacology*; D043823:Taxoids / Q000737:chemistry / Q000494:pharmacology*; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism; D050257:Tubulin Modulators / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000473:Alkanes; D000970:Antineoplastic Agents; D002219:Carbamates; D007783:Lactones; D011753:Pyrones; D043823:Taxoids; D014404:Tubulin; D050257:Tubulin Modulators; D000077143:Docetaxel; C071229:discodermolide; D017239:Paclitaxel