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Structural basis for the regulation of tubulin by vinblastine

Nature volume 435, pages 519–522 (2005)Cite this article

Abstract

Vinblastine is one of several tubulin-targeting Vinca alkaloids that have been responsible for many chemotherapeutic successes since their introduction in the clinic as antitumour drugs1. In contrast with the two other classes of small tubulin-binding molecules (Taxol2 and colchicine3), the binding site of vinblastine is largely unknown and the molecular mechanism of this drug has remained elusive. Here we report the X-ray structure of vinblastine bound to tubulin in a complex with the RB3 protein stathmin-like domain (RB3-SLD). Vinblastine introduces a wedge at the interface of two tubulin molecules and thus interferes with tubulin assembly. Together with electron microscopical and biochemical data, the structure explains vinblastine-induced tubulin self-association into spiral aggregates at the expense of microtubule growth4. It also shows that vinblastine and the amino-terminal part of RB3-SLD binding sites share a hydrophobic groove on the α-tubulin surface that is located at an intermolecular contact in microtubules. This is an attractive target for drugs designed to perturb microtubule dynamics by interfacial interference, for which tubulin seems ideally suited because of its propensity to self-associate.

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Figure 1: The vinblastine-binding site.
Figure 2: Structural changes in (Tc) 2 R after binding vinblastine.
Figure 3
Figure 4: The pseudo-first-order plot of the apparent rate constants for binding of vinblastine at the indicated concentrations to 0.3 µM (Tc) 2 R yields the values of k + (slope) and k - (ordinate intercept). The influence of SLDs on vinblastine-mediated formation of spiral aggregates.

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Acknowledgements

F.R. thanks F. Guéritte and D. Guénard for support. We thank M.-F. Carlier for help and discussions. We are grateful to the Interdisciplinary Center for Microscopy of the University of Basel for access to the transmission electron microscope. This work was supported by the Association pour la Recherche sur le Cancer, the Centre National pour la Recherche Scientifique and the Institut National pour la Santé et la Recherche Médicale. Diffraction data were collected on beamline ID14-4 at European Synchrotron Radiation Facility. C.W. was supported in part by the K.C. Wong foundation and by an ARC postdoctoral fellowship.

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Author notes

  1. Patrick A. Curmi

    Present address: Laboratoire Structure et Reconnaissance des Biomolécules, EA3637, Université Evry-Val d'Essonne, rue du Père Jarlan, 91025, Evry, France

Authors and Affiliations

  1. Laboratoire d'Enzymologie et Biochimie Structurales, UPR 9063, Centre National de la Recherche Scientifique, Bâtiment 34, 1 avenue de la Terrasse, 91198, Gif-sur-Yvette Cedex, France

    Benoît Gigant, Chunguang Wang & Marcel Knossow

  2. European Molecular Biology Laboratory (EMBL), Grenoble Outstation, 6 rue Jules Horowitz, BP 181, 38042, Grenoble, Cedex 9, France

    Raimond B. G. Ravelli

  3. Institut de Chimie des Substances Naturelles, Centre National de la Recherche Scientifique, 1 avenue de la Terrasse, 91198, Gif-sur-Yvette Cedex, France

    Fanny Roussi

  4. Paul Scherrer Institut, Biomolecular Research, Structural Biology, CH-5232, Villigen PSI, Switzerland

    Michel O. Steinmetz

  5. U706 INSERM/UPMC, Institut du Fer à Moulin, 17 rue du Fer à Moulin, 75005, Paris, France

    Patrick A. Curmi & André Sobel

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Correspondence to Marcel Knossow.

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Competing interests

Coordinates and structure factors have been deposited in the Protein Data Bank under accession number 1Z2B (tubulin–colchicine:RB3-SLD–vinblastine). Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Figure S1

Electron micrograph of negatively stained vinblastine-induced tubulin-colchicine curls. (DOC 160 kb)

Supplementary Figure S2

Interference between vinblastine and the straight tubulin assembly in protofilaments. (DOC 166 kb)

Supplementary Figure S3

The change of M-loop position after vinblastine-binding to a straight protofilament. (DOC 394 kb)

Supplementary Figure Legends

Legends to accompany the above Supplementary Figures S1-S3. (DOC 22 kb)

Supplementary Methods

Details on the proteins and buffers, structure determination and illustrations used in the study. (DOC 20 kb)

Supplementary Table S1

Crystallographic data and refinement statistics. (DOC 23 kb)

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Gigant, B., Wang, C., Ravelli, R. et al. Structural basis for the regulation of tubulin by vinblastine. Nature 435, 519–522 (2005). https://doi.org/10.1038/nature03566

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