Zampanolide, a potent new microtubule-stabilizing agent, covalently reacts with the taxane luminal site in tubulin α,β-heterodimers and microtubules

Jessica J. Field, Benet Pera, Enrique Calvo, Angeles Canales, Didier Zurwerra, Chiara Trigili, Javier Rodríguez-Salarichs, Ruth Matesanz, Arun Kanakkanthara, St John Wakefield, A. Jonathan Singh, Jesús Jiménez-Barbero, Peter Northcote, John H. Miller, Juan Antonio López, Ernest Hamel, Isabel Barasoain, Karl Heinz Altmann, José Fernando Díaz

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Zampanolide and its less active analog dactylolide compete with paclitaxel for binding to microtubules and represent a new class of microtubule-stabilizing agent (MSA). Mass spectrometry demonstrated that the mechanism of action of both compounds involved covalent binding to β-tubulin at residues N228 and H229 in the taxane site of the microtubule. Alkylation of N228 and H229 was also detected in α,β-tubulin dimers. However, unlike cyclostreptin, the other known MSA that alkylates β-tubulin, zampanolide was a strong MSA. Modeling the structure of the adducts, using the NMR-derived dactylolide conformation, indicated that the stabilizing activity of zampanolide is likely due to interactions with the M-loop. Our results strongly support the existence of the luminal taxane site of microtubules in tubulin dimers and suggest that microtubule nucleation induction by MSAs may proceed through an allosteric mechanism.

Original languageEnglish (US)
Pages (from-to)686-698
Number of pages13
JournalChemistry and Biology
Volume19
Issue number6
DOIs
StatePublished - Jun 22 2012
Externally publishedYes

Fingerprint

Excipients
Tubulin
Microtubules
Dimers
Alkylation
Paclitaxel
Mass spectrometry
Conformations
Nucleation
Nuclear magnetic resonance
zampanolide
taxane
Mass Spectrometry
dactylolide

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Cite this

Zampanolide, a potent new microtubule-stabilizing agent, covalently reacts with the taxane luminal site in tubulin α,β-heterodimers and microtubules. / Field, Jessica J.; Pera, Benet; Calvo, Enrique; Canales, Angeles; Zurwerra, Didier; Trigili, Chiara; Rodríguez-Salarichs, Javier; Matesanz, Ruth; Kanakkanthara, Arun; Wakefield, St John; Singh, A. Jonathan; Jiménez-Barbero, Jesús; Northcote, Peter; Miller, John H.; López, Juan Antonio; Hamel, Ernest; Barasoain, Isabel; Altmann, Karl Heinz; Díaz, José Fernando.

In: Chemistry and Biology, Vol. 19, No. 6, 22.06.2012, p. 686-698.

Research output: Contribution to journalArticle

Field, JJ, Pera, B, Calvo, E, Canales, A, Zurwerra, D, Trigili, C, Rodríguez-Salarichs, J, Matesanz, R, Kanakkanthara, A, Wakefield, SJ, Singh, AJ, Jiménez-Barbero, J, Northcote, P, Miller, JH, López, JA, Hamel, E, Barasoain, I, Altmann, KH & Díaz, JF 2012, 'Zampanolide, a potent new microtubule-stabilizing agent, covalently reacts with the taxane luminal site in tubulin α,β-heterodimers and microtubules', Chemistry and Biology, vol. 19, no. 6, pp. 686-698. https://doi.org/10.1016/j.chembiol.2012.05.008
Field, Jessica J. ; Pera, Benet ; Calvo, Enrique ; Canales, Angeles ; Zurwerra, Didier ; Trigili, Chiara ; Rodríguez-Salarichs, Javier ; Matesanz, Ruth ; Kanakkanthara, Arun ; Wakefield, St John ; Singh, A. Jonathan ; Jiménez-Barbero, Jesús ; Northcote, Peter ; Miller, John H. ; López, Juan Antonio ; Hamel, Ernest ; Barasoain, Isabel ; Altmann, Karl Heinz ; Díaz, José Fernando. / Zampanolide, a potent new microtubule-stabilizing agent, covalently reacts with the taxane luminal site in tubulin α,β-heterodimers and microtubules. In: Chemistry and Biology. 2012 ; Vol. 19, No. 6. pp. 686-698.
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AU - Canales, Angeles

AU - Zurwerra, Didier

AU - Trigili, Chiara

AU - Rodríguez-Salarichs, Javier

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AU - Wakefield, St John

AU - Singh, A. Jonathan

AU - Jiménez-Barbero, Jesús

AU - Northcote, Peter

AU - Miller, John H.

AU - López, Juan Antonio

AU - Hamel, Ernest

AU - Barasoain, Isabel

AU - Altmann, Karl Heinz

AU - Díaz, José Fernando

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