Substoichiometric binding of taxol suppresses microtubule dynamics.
We have measured the effects of taxol (10 nM to 1 microM) on the growing and shortening dynamics at the ends of individual bovine brain microtubules in vitro and have correlated the effects both with the stoichiometry of taxol binding to tubulin in microtubules and with the changes in the microtubule polymer mass. The results indicate that taxol suppresses microtubule dynamic instability differently depending upon the stoichiometry of taxol binding to the microtubules. At the lowest effective concentrations (< or = 100 nM), substoichiometric binding of taxol to tubulin in microtubules (between 0.001 and 0.01 mol of bound taxol/mol of tubulin in microtubules) potently and selectively suppresses the rate and extent of shortening at plus ends in association with some increase (28% to 60%) in the mass of microtubule polymer. At intermediate taxol concentrations (between 100 nM and 1 microM), the binding of additional taxol molecules to the microtubules (between 0.01 and 0.1 mol of taxol bound/mol of tubulin in microtubules) inhibits both growing and shortening events at both microtubule ends with no additional increase in microtubule polymer mass. At high taxol concentrations and high taxol binding stoichiometries (> or = 1 microM taxol and > or = 0.1 mol of taxol bound/mol of tubulin in microtubules), microtubule mass increases sharply and dynamics is almost completely suppressed. The data support the hypothesis that binding of a molecule of taxol to a tubulin subunit in microtubules induces a conformational change in that subunit that strongly reduces its ability to dissociate when the subunit becomes exposed at the microtubule end.
Division of Molecular, Cellular, Department of Biological Sciences, University of California, Santa Barbara 93106.
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Derry
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Wilson
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Jordan
D000818:Animals; D001921:Brain; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D066298:In Vitro Techniques; D007700:Kinetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism; D017239:Paclitaxel / Q000008:administration & dosage* / Q000378:metabolism; D011108:Polymers; D011485:Protein Binding / Q000187:drug effects; D014404:Tubulin / Q000378:metabolism*; D014741:Video Recording; D014747:Vinblastine / Q000378:metabolism
D008869:Microtubule-Associated Proteins; D011108:Polymers; D014404:Tubulin; D014747:Vinblastine; D017239:Paclitaxel