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Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges
Tau is a Microtubule-associated protein that induces and stabilizes the formation of the Microtubule cytoskeleton and plays an important role in neurodegenerative diseases. The Microtubules binding region of Tau has been determined for a long time but where and how Tau binds to its partner still rem...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6138654/ https://www.ncbi.nlm.nih.gov/pubmed/30218010 http://dx.doi.org/10.1038/s41598-018-32096-9 |
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author | Martinho, Marlène Allegro, Diane Huvent, Isabelle Chabaud, Charlotte Etienne, Emilien Kovacic, Hervé Guigliarelli, Bruno Peyrot, Vincent Landrieu, Isabelle Belle, Valérie Barbier, Pascale |
author_facet | Martinho, Marlène Allegro, Diane Huvent, Isabelle Chabaud, Charlotte Etienne, Emilien Kovacic, Hervé Guigliarelli, Bruno Peyrot, Vincent Landrieu, Isabelle Belle, Valérie Barbier, Pascale |
author_sort | Martinho, Marlène |
collection | PubMed |
description | Tau is a Microtubule-associated protein that induces and stabilizes the formation of the Microtubule cytoskeleton and plays an important role in neurodegenerative diseases. The Microtubules binding region of Tau has been determined for a long time but where and how Tau binds to its partner still remain a topic of debate. We used Site Directed Spin Labeling combined with EPR spectroscopy to monitor Tau upon binding to either Taxol-stabilized MTs or to αβ-tubulin when Tau is directly used as an inducer of MTs formation. Using maleimide-functionalized labels grafted on the two natural cysteine residues of Tau, we found in both cases that Tau remains highly flexible in these regions confirming the fuzziness of Tau:MTs complexes. More interestingly, using labels linked by a disulfide bridge, we evidenced for the first time thiol disulfide exchanges between αβ-tubulin or MTs and Tau. Additionally, Tau fragments having the two natural cysteines or variants containing only one of them were used to determine the role of each cysteine individually. The difference observed in the label release kinetics between preformed MTs or Tau-induced MTs, associated to a comparison of structural data, led us to propose two putative binding sites of Tau on αβ-tubulin. |
format | Online Article Text |
id | pubmed-6138654 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-61386542018-09-15 Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges Martinho, Marlène Allegro, Diane Huvent, Isabelle Chabaud, Charlotte Etienne, Emilien Kovacic, Hervé Guigliarelli, Bruno Peyrot, Vincent Landrieu, Isabelle Belle, Valérie Barbier, Pascale Sci Rep Article Tau is a Microtubule-associated protein that induces and stabilizes the formation of the Microtubule cytoskeleton and plays an important role in neurodegenerative diseases. The Microtubules binding region of Tau has been determined for a long time but where and how Tau binds to its partner still remain a topic of debate. We used Site Directed Spin Labeling combined with EPR spectroscopy to monitor Tau upon binding to either Taxol-stabilized MTs or to αβ-tubulin when Tau is directly used as an inducer of MTs formation. Using maleimide-functionalized labels grafted on the two natural cysteine residues of Tau, we found in both cases that Tau remains highly flexible in these regions confirming the fuzziness of Tau:MTs complexes. More interestingly, using labels linked by a disulfide bridge, we evidenced for the first time thiol disulfide exchanges between αβ-tubulin or MTs and Tau. Additionally, Tau fragments having the two natural cysteines or variants containing only one of them were used to determine the role of each cysteine individually. The difference observed in the label release kinetics between preformed MTs or Tau-induced MTs, associated to a comparison of structural data, led us to propose two putative binding sites of Tau on αβ-tubulin. Nature Publishing Group UK 2018-09-14 /pmc/articles/PMC6138654/ /pubmed/30218010 http://dx.doi.org/10.1038/s41598-018-32096-9 Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Martinho, Marlène Allegro, Diane Huvent, Isabelle Chabaud, Charlotte Etienne, Emilien Kovacic, Hervé Guigliarelli, Bruno Peyrot, Vincent Landrieu, Isabelle Belle, Valérie Barbier, Pascale Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title | Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title_full | Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title_fullStr | Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title_full_unstemmed | Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title_short | Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges |
title_sort | two tau binding sites on tubulin revealed by thiol-disulfide exchanges |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6138654/ https://www.ncbi.nlm.nih.gov/pubmed/30218010 http://dx.doi.org/10.1038/s41598-018-32096-9 |
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