Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges

Abstract 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...

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Autores principales:	Marlène Martinho, Diane Allegro, Isabelle Huvent, Charlotte Chabaud, Emilien Etienne, Hervé Kovacic, Bruno Guigliarelli, Vincent Peyrot, Isabelle Landrieu, Valérie Belle, Pascale Barbier
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Thiol-disulfide Exchange Label Release Native Cysteine Residues Spin Label Co-sedimentation Assay Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f38fd31e0683461fb98ee95d22a0e5fa
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spelling	oai:doaj.org-article:f38fd31e0683461fb98ee95d22a0e5fa2021-12-02T15:09:03ZTwo Tau binding sites on tubulin revealed by thiol-disulfide exchanges10.1038/s41598-018-32096-92045-2322https://doaj.org/article/f38fd31e0683461fb98ee95d22a0e5fa2018-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-32096-9https://doaj.org/toc/2045-2322Abstract 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.Marlène MartinhoDiane AllegroIsabelle HuventCharlotte ChabaudEmilien EtienneHervé KovacicBruno GuigliarelliVincent PeyrotIsabelle LandrieuValérie BellePascale BarbierNature PortfolioarticleThiol-disulfide ExchangeLabel ReleaseNative Cysteine ResiduesSpin LabelCo-sedimentation AssayMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Thiol-disulfide Exchange Label Release Native Cysteine Residues Spin Label Co-sedimentation Assay Medicine R Science Q
spellingShingle	Thiol-disulfide Exchange Label Release Native Cysteine Residues Spin Label Co-sedimentation Assay Medicine R Science Q Marlène Martinho Diane Allegro Isabelle Huvent Charlotte Chabaud Emilien Etienne Hervé Kovacic Bruno Guigliarelli Vincent Peyrot Isabelle Landrieu Valérie Belle Pascale Barbier Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges
description	Abstract 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	article
author	Marlène Martinho Diane Allegro Isabelle Huvent Charlotte Chabaud Emilien Etienne Hervé Kovacic Bruno Guigliarelli Vincent Peyrot Isabelle Landrieu Valérie Belle Pascale Barbier
author_facet	Marlène Martinho Diane Allegro Isabelle Huvent Charlotte Chabaud Emilien Etienne Hervé Kovacic Bruno Guigliarelli Vincent Peyrot Isabelle Landrieu Valérie Belle Pascale Barbier
author_sort	Marlène Martinho
title	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_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_sort	two tau binding sites on tubulin revealed by thiol-disulfide exchanges
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/f38fd31e0683461fb98ee95d22a0e5fa
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Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges

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