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Structural characterization by nuclear magnetic resonance of the impact of phosphorylation in the proline-rich region of the disordered Tau protein

Authors

  • Nathalie Sibille,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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    • Nathalie Sibille and Isabelle Huvent contributed equally to this work.

  • Isabelle Huvent,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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    • Nathalie Sibille and Isabelle Huvent contributed equally to this work.

  • Caroline Fauquant,

    1. Laboratoire d'Enzymologie et Biochimie Structurales, Centre de Recherche de Gif, CNRS, 91198 Gif-sur-Yvette, France
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  • Dries Verdegem,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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  • Laziza Amniai,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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  • Arnaud Leroy,

    1. Laboratoire de Biochimie Appliquée, Faculté de Pharmacie (Paris XI), Chatenay-Malabry, France
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  • Jean-Michel Wieruszeski,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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  • Guy Lippens,

    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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  • Isabelle Landrieu

    Corresponding author
    1. CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
    • CNRS-UMR 8576 UGSF-IFR 147, Université des Sciences et Technologies de Lille 1, 59655 Villeneuve d'Ascq Cedex, France
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  • Backbone and Cβ chemical shift assignments of TauF4 have been deposited at BMRB (www.bmrb.wisc.edu) under accession number 17945.

Abstract

Phosphorylation of the neuronal Tau protein is implicated in both the regulation of its physiological function of microtubule stabilization and its pathological propensity to aggregate into the fibers that characterize Alzheimer's diseased neurons. However, how specific phosphorylation events influence both aspects of Tau biology remains largely unknown. In this study, we address the structural impact of phosphorylation of the Tau protein by Nuclear Magnetic Resonance (NMR) spectroscopy on a functional fragment of Tau (Tau[Ser208–Ser324] = TauF4). TauF4 was phosphorylated by the proline-directed CDK2/CycA3 kinase on Thr231 (generating the AT180 epitope), Ser235, and equally on Thr212 and Thr217 in the Proline-rich region (Tau[Ser208-Gln244] or PRR). These modifications strongly decrease the capacity of TauF4 to polymerize tubulin into microtubules. While all the NMR parameters are consistent with a globally disordered Tau protein fragment, local clusters of structuration can be defined. The most salient result of our NMR analysis is that phosphorylation in the PRR stabilizes a short α-helix that runs from pSer235 till the very beginning of the microtubule-binding region (Tau[Thr245-Ser324] or MTBR of TauF4). Phosphorylation of Thr231/Ser235 creates a N-cap with helix stabilizing role while phosphorylation of Thr212/Thr217 does not induce modification of the local transient secondary structure, showing that the stabilizing effect is sequence specific. Using paramagnetic relaxation experiments, we additionally show a transient interaction between the PRR and the MTBR, observed in both TauF4 and phospho-TauF4. Proteins 2012. © 2011 Wiley Periodicals, Inc.

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