Home LiteratureArticle Details
PMID: 18405918 Published · ppublish English Journal Article Research Support, N.I.H., Intramural Research Support, Non-U.S. Gov't

A unique mode of microtubule stabilization induced by peloruside A.

Journal of molecular biology ·Vol. 378 ·No. 5 ·2008-05-16 ·Pages 1016-30

Huzil JT, Chik JK, Slysz GW, Freedman H, Tuszynski J, Taylor RE, Sackett DL, Schriemer DC

Abstract

Microtubules are significant therapeutic targets for the treatment of cancer, where suppression of microtubule dynamicity by drugs such as paclitaxel forms the basis of clinical efficacy. Peloruside A, a macrolide isolated from New Zealand marine sponge Mycale hentscheli, is a microtubule-stabilizing agent that synergizes with taxoid drugs through a unique site and is an attractive lead compound in the development of combination therapies. We report here unique allosteric properties of microtubule stabilization via peloruside A and present a structural model of the peloruside-binding site. Using a strategy involving comparative hydrogen-deuterium exchange mass spectrometry of different microtubule-stabilizing agents, we suggest that taxoid-site ligands epothilone A and docetaxel stabilize microtubules primarily through improved longitudinal interactions centered on the interdimer interface, with no observable contributions from lateral interactions between protofilaments. The mode by which peloruside A achieves microtubule stabilization also involves the interdimer interface, but includes contributions from the alpha/beta-tubulin intradimer interface and protofilament contacts, both in the form of destabilizations. Using data-directed molecular docking simulations, we propose that peloruside A binds within a pocket on the exterior of beta-tubulin at a previously unknown ligand site, rather than on alpha-tubulin as suggested in earlier studies.

MeSH Terms
Allosteric Regulation Amino Acid Sequence Animals Antineoplastic Agents, Phytogenic/chemistry,metabolism Binding Sites Bridged Bicyclo Compounds, Heterocyclic/chemistry,metabolism Cattle Dimerization Guanosine Triphosphate/analogs & derivatives,chemistry Lactones/chemistry,metabolism Ligands Mass Spectrometry Microtubules/chemistry,metabolism Models, Molecular Molecular Sequence Data Peptides/chemistry,genetics,metabolism Protein Isoforms/chemistry,genetics,metabolism Protein Structure, Quaternary Sequence Alignment Tubulin/chemistry,genetics,metabolism Tubulin Modulators/chemistry,metabolism
Chemicals
Antineoplastic Agents, Phytogenic Bridged Bicyclo Compounds, Heterocyclic Lactones Ligands Peptides Protein Isoforms Tubulin Tubulin Modulators peloruside A 5'-guanylylmethylenebisphosphonate Guanosine Triphosphate
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Huzil J Torin
Division of Experimental Oncology, Cross Cancer Institute, 11560 University Avenue, Edmonton, Alberta, Canada.
Chik John K
Slysz Gordon W
Freedman Holly
Tuszynski Jack
Taylor Richard E
Sackett Dan L
Schriemer David C
References (49)
49 references, click to expand
  1. Microtubule stabilizing agents: their molecular signaling consequences and the potential for enhancement by drug combination.
    Cancer Treat Rev. 2006 May;32(3):166-79 PMID: 16527420
  2. Refined structure of alpha beta-tubulin at 3.5 A resolution.
    J Mol Biol. 2001 Nov 9;313(5):1045-57 PMID: 11700061
  3. Hydrogen/deuterium-exchange (H/D-Ex) of PPARgamma LBD in the presence of various modulators.
    Protein Sci. 2006 Aug;15(8):1883-92 PMID: 16823031
  4. Paclitaxel in a novel formulation containing less Cremophor EL as first-line therapy for advanced breast cancer: a phase II trial.
    Invest New Drugs. 2005 Mar;23(2):171-7 PMID: 15744594
  5. Characterization of microtubule protofilament numbers. How does the surface lattice accommodate?
    J Mol Biol. 1990 Apr 20;212(4):775-86 PMID: 2329582
  6. Computational comparison of microtubule-stabilising agents laulimalide and peloruside with taxol and colchicine.
    Bioorg Med Chem Lett. 2004 Oct 4;14(19):4825-9 PMID: 15341932
  7. NMR determination of the bioactive conformation of peloruside A bound to microtubules.
    J Am Chem Soc. 2006 Jul 12;128(27):8757-65 PMID: 16819869
  8. Role of GTP hydrolysis in microtubule dynamics: information from a slowly hydrolyzable analogue, GMPCPP.
    Mol Biol Cell. 1992 Oct;3(10):1155-67 PMID: 1421572
  9. Taxol differentially modulates the dynamics of microtubules assembled from unfractionated and purified beta-tubulin isotypes.
    Biochemistry. 1997 Mar 25;36(12):3554-62 PMID: 9132006
  10. Identification of the protein kinase A regulatory RIalpha-catalytic subunit interface by amide H/2H exchange and protein docking.
    Proc Natl Acad Sci U S A. 2003 Nov 11;100(23):13264-9 PMID: 14583592
  11. Hydrogen exchange/electrospray ionization mass spectrometry studies of structural features of proteins and protein/protein interactions.
    Anal Biochem. 1999 Feb 15;267(2):252-9 PMID: 10036128
  12. The betaI/betaIII-tubulin isoforms and their complexes with antimitotic agents. Docking and molecular dynamics studies.
    FEBS J. 2006 Jul;273(14):3301-10 PMID: 16803461
  13. Hypersensitivity reactions from taxol.
    J Clin Oncol. 1990 Jul;8(7):1263-8 PMID: 1972736
  14. Hydrogen/deuterium exchange mass spectrometry of actin in various biochemical contexts.
    J Mol Biol. 2003 Nov 28;334(3):373-85 PMID: 14623181
  15. Preparation of a monoclonal antibody specific for the class IV isotype of beta-tubulin. Purification and assembly of alpha beta II, alpha beta III, and alpha beta IV tubulin dimers from bovine brain.
    J Biol Chem. 1992 Mar 15;267(8):5625-30 PMID: 1544937
  16. Peloruside A does not bind to the taxoid site on beta-tubulin and retains its activity in multidrug-resistant cell lines.
    Cancer Res. 2004 Aug 1;64(15):5063-7 PMID: 15289305
  17. Microtubule interactions with chemically diverse stabilizing agents: thermodynamics of binding to the paclitaxel site predicts cytotoxicity.
    Chem Biol. 2005 Dec;12(12):1269-79 PMID: 16356844
  18. Conformations of laulimalide in DMSO-d6.
    J Am Chem Soc. 2005 Sep 21;127(37):12838-46 PMID: 16159277
  19. Peloruside A, a novel antimitotic agent with paclitaxel-like microtubule- stabilizing activity.
    Cancer Res. 2002 Jun 15;62(12):3356-60 PMID: 12067973
  20. The Amber biomolecular simulation programs.
    J Comput Chem. 2005 Dec;26(16):1668-88 PMID: 16200636
  21. The binding mode of epothilone A on alpha,beta-tubulin by electron crystallography.
    Science. 2004 Aug 6;305(5685):866-9 PMID: 15297674
  22. Changes in microtubule protofilament number induced by Taxol binding to an easily accessible site. Internal microtubule dynamics.
    J Biol Chem. 1998 Dec 11;273(50):33803-10 PMID: 9837970
  23. Peloruside A: a potent cytotoxic macrolide isolated from the new zealand marine sponge Mycale sp.
    J Org Chem. 2000 Jan 28;65(2):445-9 PMID: 10813954
  24. Peloruside A synergizes with other microtubule stabilizing agents in cultured cancer cell lines.
    Mol Pharm. 2007 Mar-Apr;4(2):269-80 PMID: 17397239
  25. PRODRG: a tool for high-throughput crystallography of protein-ligand complexes.
    Acta Crystallogr D Biol Crystallogr. 2004 Aug;60(Pt 8):1355-63 PMID: 15272157
  26. Blind docking of drug-sized compounds to proteins with up to a thousand residues.
    FEBS Lett. 2006 Feb 20;580(5):1447-50 PMID: 16460734
  27. Insights into the mechanism of microtubule stabilization by Taxol.
    Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10166-10173 PMID: 16801540
  28. High-resolution model of the microtubule.
    Cell. 1999 Jan 8;96(1):79-88 PMID: 9989499
  29. Hydrogen exchange methods to study protein folding.
    Methods. 2004 Sep;34(1):51-64 PMID: 15283915
  30. Structural basis of the activity of the microtubule-stabilizing agent epothilone a studied by NMR spectroscopy in solution.
    Angew Chem Int Ed Engl. 2007;46(11):1864-8 PMID: 17274084
  31. Understanding noncovalent interactions: ligand binding energy and catalytic efficiency from ligand-induced reductions in motion within receptors and enzymes.
    Angew Chem Int Ed Engl. 2004 Dec 10;43(48):6596-616 PMID: 15593167
  32. Assembly of GMPCPP-bound tubulin into helical ribbons and tubes and effect of colchicine.
    Cell Cycle. 2005 Sep;4(9):1157-60 PMID: 16123589
  33. Structural mechanisms underlying nucleotide-dependent self-assembly of tubulin and its relatives.
    Curr Opin Struct Biol. 2006 Apr;16(2):221-9 PMID: 16549346
  34. Suppression of microtubule dynamic instability and treadmilling by deuterium oxide.
    Biochemistry. 2000 May 2;39(17):5075-81 PMID: 10819973
  35. The microtubule stabilizing agent laulimalide does not bind in the taxoid site, kills cells resistant to paclitaxel and epothilones, and may not require its epoxide moiety for activity.
    Biochemistry. 2002 Jul 23;41(29):9109-15 PMID: 12119025
  36. Cremophor EL: the drawbacks and advantages of vehicle selection for drug formulation.
    Eur J Cancer. 2001 Sep;37(13):1590-8 PMID: 11527683
  37. Mutations in beta-tubulin map to domains involved in regulation of microtubule stability in epothilone-resistant cell lines.
    Mol Cancer Ther. 2001 Nov;1(1):3-10 PMID: 12467233
  38. Protein conformations, interactions, and H/D exchange.
    Methods Enzymol. 2005;402:312-60 PMID: 16401514
  39. Microtubule polymerization dynamics.
    Annu Rev Cell Dev Biol. 1997;13:83-117 PMID: 9442869
  40. Synergistic effects of peloruside A and laulimalide with taxoid site drugs, but not with each other, on tubulin assembly.
    Mol Pharmacol. 2006 Nov;70(5):1555-64 PMID: 16887932
  41. Laulimalide and isolaulimalide, new paclitaxel-like microtubule-stabilizing agents.
    Cancer Res. 1999 Feb 1;59(3):653-60 PMID: 9973214
  42. Assembly of purified GDP-tubulin into microtubules induced by taxol and taxotere: reversibility, ligand stoichiometry, and competition.
    Biochemistry. 1993 Mar 23;32(11):2747-55 PMID: 8096151
  43. Total synthesis of (+)-peloruside A.
    Org Lett. 2005 Mar 31;7(7):1303-5 PMID: 15787492
  44. Physiochemical aspects of tubulin-interacting antimitotic drugs.
    Curr Pharm Des. 2001 Sep;7(13):1213-28 PMID: 11472263
  45. Automated docking of maltose, 2-deoxymaltose, and maltotetraose into the soybean beta-amylase active site.
    Proteins. 1999 Nov 1;37(2):166-75 PMID: 10584063
  46. Nucleotide-dependent bending flexibility of tubulin regulates microtubule assembly.
    Nature. 2005 Jun 16;435(7044):911-5 PMID: 15959508
  47. Laulimalide and paclitaxel: a comparison of their effects on tubulin assembly and their synergistic action when present simultaneously.
    Mol Pharmacol. 2004 Jul;66(1):113-21 PMID: 15213302
  48. Protein hydrogen exchange mechanism: local fluctuations.
    Protein Sci. 2003 Jan;12(1):153-60 PMID: 12493838
  49. Probing protein ligand interactions by automated hydrogen/deuterium exchange mass spectrometry.
    Anal Chem. 2006 Feb 15;78(4):1005-14 PMID: 16478090
Article Info
Journal
Journal of molecular biology
Abbr.
J Mol Biol
ISSN
1089-8638
DOI
10.1016/j.jmb.2008.03.026
Published
2008-05-16
Epub
2008-00-19
Pages
1016-30
Language
English
Region
England
NLM ID
2985088R
PMCID
PMC2441936
Subset
IM
Grants
NIGMS NIH HHS · R01 GM077683 · United States
NIGMS NIH HHS · R01 GM077683-06 · United States
NIGMS NIH HHS · R01 GM077683-07 · United States
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1016/j.jmb.2008.03.026 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2