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

Marlène Martinho 1 Diane Allegro 2 Isabelle Huvent 3 Charlotte Chabaud Emilien Etienne 1 Hervé Kovacic 2 Bruno Guigliarelli 1 Vincent Peyrot 2 Isabelle Landrieu 3 Valérie Belle 1 Pascale Barbier 2
1 BIP - Bioénergétique et Ingénierie des Protéines
2 INP - Institut de neurophysiopathologie
3 UGSF - Unité de Glycobiologie Structurale et Fonctionnelle - UMR 8576
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.
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Journal articles
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Contributor : Ludovic Leloup <>
Submitted on : Friday, September 28, 2018 - 6:00:57 PM
Last modification on : Thursday, October 4, 2018 - 1:49:52 PM

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Marlène Martinho, Diane Allegro, Isabelle Huvent, Charlotte Chabaud, Emilien Etienne, et al.. Two Tau binding sites on tubulin revealed by thiol-disulfide exchanges. Scientific Reports, Nature Publishing Group, 2018, 8 (1), pp.13846. ⟨10.1038/s41598-018-32096-9⟩. ⟨hal-01883844⟩

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