Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization

Marina Khrapunovich-Baine, Vilas Menon, Pascal Verdier-Pinard, Amos B. Smith, Ruth Hogue Angeletti, Andras Fiser, Susan Band Horwitz, Hui Xiao

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The microtubule cytoskeleton has proven to be an effective target for cancer therapeutics. One class of drugs, known as microtubule stabilizing agents (MSAs), binds to microtubule polymers and stabilizes them against depolymerization. The prototype of this group of drugs, Taxol, is an effective chemotherapeutic agent used extensively in the treatment of human ovarian, breast, and lung carcinomas. Although electron crystallography and photoaffinity labeling experiments determined that the binding site for Taxol is in a hydrophobic pocket in β-tubulin, little was known about the effects of this drug on the conformation of the entire microtubule. A recent study from our laboratory utilizing hydrogen-deuterium exchange (HDX) in concert with various mass spectrometry (MS) techniques has provided new information on the structure of microtubules upon Taxol binding. In the current study we apply this technique to determine the binding mode and the conformational effects on chicken erythrocyte tubulin (CET) of anotherMSA, discodermolide, whose synthetic analogues may have potential use in the clinic. We confirmed that, like Taxol, discodermolide binds to the taxane binding pocket in β-tubulin. However, as opposed to Taxol, which has major interactions with the M-loop, discodermolide orients itself away from this loop and toward the N-terminal H1-S2 loop. Additionally, discodermolide stabilizes microtubules mainly via its effects on interdimer contacts, specifically on the α-tubulin side, and to a lesser extent on interprotofilament contacts between adjacent β-tubulin subunits. Also, our results indicate complementary stabilizing effects of Taxol and discodermolide on the microtubules, which may explain the synergy observed between the two drugs in vivo.

Original languageEnglish (US)
Pages (from-to)11664-11677
Number of pages14
JournalBiochemistry
Volume48
Issue number49
DOIs
StatePublished - Dec 15 2009

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Paclitaxel
Microtubules
Tubulin
Stabilization
Pharmaceutical Preparations
Depolymerization
Crystallography
Deuterium
Excipients
Labeling
Mass spectrometry
Conformations
discodermolide
Cytoskeleton
Hydrogen
Polymers
Binding Sites
Chickens
Mass Spectrometry
Erythrocytes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Khrapunovich-Baine, M., Menon, V., Verdier-Pinard, P., Smith, A. B., Angeletti, R. H., Fiser, A., ... Xiao, H. (2009). Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization. Biochemistry, 48(49), 11664-11677. https://doi.org/10.1021/bi901351q

Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization. / Khrapunovich-Baine, Marina; Menon, Vilas; Verdier-Pinard, Pascal; Smith, Amos B.; Angeletti, Ruth Hogue; Fiser, Andras; Band Horwitz, Susan; Xiao, Hui.

In: Biochemistry, Vol. 48, No. 49, 15.12.2009, p. 11664-11677.

Research output: Contribution to journalArticle

Khrapunovich-Baine, M, Menon, V, Verdier-Pinard, P, Smith, AB, Angeletti, RH, Fiser, A, Band Horwitz, S & Xiao, H 2009, 'Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization', Biochemistry, vol. 48, no. 49, pp. 11664-11677. https://doi.org/10.1021/bi901351q
Khrapunovich-Baine M, Menon V, Verdier-Pinard P, Smith AB, Angeletti RH, Fiser A et al. Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization. Biochemistry. 2009 Dec 15;48(49):11664-11677. https://doi.org/10.1021/bi901351q
Khrapunovich-Baine, Marina ; Menon, Vilas ; Verdier-Pinard, Pascal ; Smith, Amos B. ; Angeletti, Ruth Hogue ; Fiser, Andras ; Band Horwitz, Susan ; Xiao, Hui. / Distinct pose of discodermolide in taxol binding pocket drives a complementary mode of microtubule stabilization. In: Biochemistry. 2009 ; Vol. 48, No. 49. pp. 11664-11677.
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