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In silico design of tubulin-targeted antimitotic peptides

Nature Chemistry volume 1, pages 642–648 (2009)Cite this article

Abstract

Microtubules are polymeric structures formed by the self-assembly of tubulin dimers. The growth and shrinkage of these dynamic arrays have a key role during the cell-proliferation process. This makes tubulin the molecular target of many anticancer drugs currently in use or under clinical trial. Their impressive success is limited by the onset of resistant tumour cells during the treatment, so new resistance-proof molecules need to be developed. Here we use molecular dynamics and free-energy calculations to study the network of interactions that allow microtubule formation. Modelling the protein–protein interface allows us to identify the amino acids responsible for tubulin–tubulin binding and thus to design peptides, which correspond to tubulin subsequences, that interfere with microtubule formation. We show that the application of molecular modelling techniques leads to the identification of peptides that exhibit antitubulin activity both in vitro and in cultured cells.

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Figure 1: PPIs in tubulin dimers.
Figure 2: Structural modifications that occur in two tubulin subsequences.
Figure 3: Comparison of alanine scanning of residues that correspond to the three peptides selected at the interface.
Figure 4: The influence of peptides on tubulin assembly in vitro.
Figure 5: Microtubule organization in human lung carcinoma cell line A549 exposed for 24 hours to different treatments.

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Acknowledgements

S. Rendine is acknowledged for useful discussions.

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Authors and Affiliations

  1. Dipartimento di Chimica Fisica ed Elettrochimica, Università degli Studi di Milano, Via Golgi 19, Milano, Italy

    Stefano Pieraccini, Giorgio Saladino & Maurizio Sironi

  2. Dipartimento di Biologia, Università degli Studi di Milano, Via Celoria 26, Milano, Italy

    Graziella Cappelletti & Daniele Cartelli

  3. Dipartimento di Chimica Organica ed Industriale, Università degli Studi di Milano, Via Venezian 21, Milano, Italy

    Pierangelo Francescato, Giovanna Speranza & Paolo Manitto

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  1. Stefano Pieraccini
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  2. Giorgio Saladino
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  8. Maurizio Sironi
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Contributions

S.P. and M.S. conceived and designed the research project; G.Sa. G.C., D.C., P.F., G.Sp. and P.M. performed the experiments; S.P., G.Sa., G.C. and M.S. analysed the data and co-wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to Maurizio Sironi.

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Pieraccini, S., Saladino, G., Cappelletti, G. et al. In silico design of tubulin-targeted antimitotic peptides. Nature Chem 1, 642–648 (2009). https://doi.org/10.1038/nchem.401

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