Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands
It has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, inclu...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
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MDPI AG
2019-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/20/6/1392 |
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| author | Francisco de Asís Balaguer Tobias Mühlethaler Juan Estévez-Gallego Enrique Calvo Juan Francisco Giménez-Abián April L. Risinger Erik J. Sorensen Christopher D. Vanderwal Karl-Heinz Altmann Susan L. Mooberry Michel O. Steinmetz María Ángela Oliva Andrea E. Prota J. Fernando Díaz |
| author_facet | Francisco de Asís Balaguer Tobias Mühlethaler Juan Estévez-Gallego Enrique Calvo Juan Francisco Giménez-Abián April L. Risinger Erik J. Sorensen Christopher D. Vanderwal Karl-Heinz Altmann Susan L. Mooberry Michel O. Steinmetz María Ángela Oliva Andrea E. Prota J. Fernando Díaz |
| author_sort | Francisco de Asís Balaguer |
| collection | DOAJ |
| description | It has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, including cyclostreptin, to gain further insight into this mechanism. The crystal structure of cyclostreptin-bound tubulin reveals covalent binding to βHis229, but no stabilization of the M-loop. The capacity of cyclostreptin to induce microtubule assembly compared to other covalent taxane-site agents demonstrates that the induction of tubulin assembly is not strictly dependent on M-loop stabilization. We further demonstrate that most covalent taxane-site ligands are able to partially overcome drug resistance mediated by βIII-tubulin (βIII) overexpression in HeLa cells, and compare their activities to pironetin, an interfacial covalent inhibitor of tubulin assembly that displays invariant growth inhibition in these cells. Our findings suggest a relationship between a diminished interaction of taxane-site ligands with βIII-tubulin and βIII tubulin-mediated drug resistance. This supports the idea that overexpression of βIII increases microtubule dynamicity by counteracting the enhanced microtubule stability promoted by covalent taxane-site binding ligands. |
| first_indexed | 2024-04-13T19:14:56Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-13T19:14:56Z |
| publishDate | 2019-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-abae6ee18f044a3685a5886a0f71911a2022-12-22T02:33:42ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-03-01206139210.3390/ijms20061392ijms20061392Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site LigandsFrancisco de Asís Balaguer0Tobias Mühlethaler1Juan Estévez-Gallego2Enrique Calvo3Juan Francisco Giménez-Abián4April L. Risinger5Erik J. Sorensen6Christopher D. Vanderwal7Karl-Heinz Altmann8Susan L. Mooberry9Michel O. Steinmetz10María Ángela Oliva11Andrea E. Prota12J. Fernando Díaz13Structural and Chemical Biology Department. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, SpainLaboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232 Villigen PSI, SwitzerlandStructural and Chemical Biology Department. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, SpainUnidad de Proteómica. Centro Nacional de Investigaciones Cardiovasculares, CNIC. Melchor Fernández de Almagro 3, 28029 Madrid, SpainStructural and Chemical Biology Department. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, SpainDepartment of Pharmacology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USADepartment of Chemistry, Princeton University, Princeton, NJ 08544, USADepartment of Chemistry, 1102 Natural Sciences II, University of California, Irvine, CA 92697-2025, USAETH Zürich, Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, 8093 Zürich, SwitzerlandDepartment of Pharmacology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USALaboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232 Villigen PSI, SwitzerlandStructural and Chemical Biology Department. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, SpainLaboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, 5232 Villigen PSI, SwitzerlandStructural and Chemical Biology Department. Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, SpainIt has been proposed that one of the mechanisms of taxane-site ligand-mediated tubulin activation is modulation of the structure of a switch element (the M-loop) from a disordered form in dimeric tubulin to a folded helical structure in microtubules. Here, we used covalent taxane-site ligands, including cyclostreptin, to gain further insight into this mechanism. The crystal structure of cyclostreptin-bound tubulin reveals covalent binding to βHis229, but no stabilization of the M-loop. The capacity of cyclostreptin to induce microtubule assembly compared to other covalent taxane-site agents demonstrates that the induction of tubulin assembly is not strictly dependent on M-loop stabilization. We further demonstrate that most covalent taxane-site ligands are able to partially overcome drug resistance mediated by βIII-tubulin (βIII) overexpression in HeLa cells, and compare their activities to pironetin, an interfacial covalent inhibitor of tubulin assembly that displays invariant growth inhibition in these cells. Our findings suggest a relationship between a diminished interaction of taxane-site ligands with βIII-tubulin and βIII tubulin-mediated drug resistance. This supports the idea that overexpression of βIII increases microtubule dynamicity by counteracting the enhanced microtubule stability promoted by covalent taxane-site binding ligands.https://www.mdpi.com/1422-0067/20/6/1392cyclostreptintubulinmicrotubulesmultidrug resistancetaxanes |
| spellingShingle | Francisco de Asís Balaguer Tobias Mühlethaler Juan Estévez-Gallego Enrique Calvo Juan Francisco Giménez-Abián April L. Risinger Erik J. Sorensen Christopher D. Vanderwal Karl-Heinz Altmann Susan L. Mooberry Michel O. Steinmetz María Ángela Oliva Andrea E. Prota J. Fernando Díaz Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands International Journal of Molecular Sciences cyclostreptin tubulin microtubules multidrug resistance taxanes |
| title | Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands |
| title_full | Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands |
| title_fullStr | Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands |
| title_full_unstemmed | Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands |
| title_short | Crystal Structure of the Cyclostreptin-Tubulin Adduct: Implications for Tubulin Activation by Taxane-Site Ligands |
| title_sort | crystal structure of the cyclostreptin tubulin adduct implications for tubulin activation by taxane site ligands |
| topic | cyclostreptin tubulin microtubules multidrug resistance taxanes |
| url | https://www.mdpi.com/1422-0067/20/6/1392 |
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