A designed ankyrin repeat protein selected to bind to tubulin caps the microtubule plus end
| Autor: | Andreas Plückthun, Christian Duellberg, Ludovic Pecqueur, Birgit Dreier, Benoît Gigant, Qiyang Jiang, Thomas Surrey, Marcel Knossow, Chunguang Wang |
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| Přispěvatelé: | Department of Biochemistry [Zurich], Universität Zürich [Zürich] = University of Zurich (UZH), Laboratoire d’Enzymologie et Biochimie Structurales, Centre de Recherche de Gif, Centre National de la Recherche Scientifique, 91198 Gif sur Yvette, France, Laboratoire d'Enzymologie et Biochimie Structurales, Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), University of Zurich, Knossow, M |
| Rok vydání: | 2012 |
| Předmět: |
Models
Molecular Xenopus Fluorescence Polarization [SDV.BC]Life Sciences [q-bio]/Cellular Biology macromolecular substances Crystallography X-Ray Protein Engineering Microtubules Microtubule polymerization 03 medical and health sciences 0302 clinical medicine Tubulin Microtubule 10019 Department of Biochemistry Animals Microtubule end [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] Cytoskeleton DNA Primers 030304 developmental biology Microtubule nucleation 1000 Multidisciplinary 0303 health sciences Multidisciplinary [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] biology Biological Sciences Ankyrin Repeat Cell biology Microtubule plus-end Microscopy Fluorescence DARPin Multiprotein Complexes biology.protein 570 Life sciences Guanosine Triphosphate 030217 neurology & neurosurgery |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America Proceedings of the National Academy of Sciences of the United States of America, National Academy of Sciences, 2012, 109 (30), pp.12011-12016. ⟨10.1073/pnas.1204129109⟩ |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.1204129109 |
| Popis: | International audience; Microtubules are cytoskeleton filaments consisting of αβ-tubulin heterodimers. They switch between phases of growth and shrinkage. The underlying mechanism of this property, called dynamic instability, is not fully understood. Here, we identified a designed ankyrin repeat protein (DARPin) that interferes with microtubule assembly in a unique manner. The X-ray structure of its complex with GTP-tubulin shows that it binds to the β-tubulin surface exposed at microtubule (+) ends. The details of the structure provide insight into the role of GTP in microtubule polymerization and the conformational state of tubulin at the very microtubule end. They show in particular that GTP facilitates the tubulin structural switch that accompanies microtubule assembly but does not trigger it in unpolymerized tubulin. Total internal reflection fluorescence microscopy revealed that the DARPin specifically blocks growth at the microtubule (+) end by a selective end-capping mechanism, ultimately favoring microtubule disassembly from that end. DARPins promise to become designable tools for the dissection of microtubule dynamic properties selective for either of their two different ends. |
| Databáze: | OpenAIRE |
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