Phosphorylated full-length Tau interacts with 14-3-3 proteins via two short phosphorylated sequences, each occupying a binding groove of 14-3-3 dimer

João Filipe Neves, Olivia Petrvalská, Francesco Bosica, François Xavier Cantrelle, Hamida Merzougui, Gavin O'Mahony, Xavier Hanoulle, Tomáš Obšil, Isabelle Landrieu (Corresponding author)

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11 Citations (Scopus)

Abstract

Protein–protein interactions (PPIs) remain poorly explored targets for the treatment of Alzheimer’s disease. The interaction of 14-3-3 proteins with Tau was shown to be linked to Tau pathology. This PPI is therefore seen as a potential target for Alzheimer’s disease. When Tau is phosphorylated by PKA (Tau-PKA), several phosphorylation sites are generated, including two known 14-3-3 binding sites, surrounding the phosphorylated serines 214 and 324 of Tau. The crystal structures of 14-3-3 in complex with peptides surrounding these Tau phosphosites show that both these motifs are anchored in the amphipathic binding groove of 14-3-3. However, in the absence of structural data with the full-length Tau protein, the stoichiometry of the complex or the interface and affinity of the partners is still unclear. In this work, we addressed these points, using a broad range of biophysical techniques. The interaction of the long and disordered Tau-PKA protein with 14-3-3σ is restricted to two short sequences, containing phosphorylated serines, which bind in the amphipathic binding groove of 14-3-3σ. Phosphorylation of Tau is fundamental for the formation of this stable complex, and the affinity of the Tau-PKA/14-3-3σ interaction is in the 1–10 micromolar range. Each monomer of the 14-3-3σ dimer binds one of two different phosphorylated peptides of Tau-PKA, suggesting a 14-3-3/Tau-PKA stoichiometry of 2 : 1, confirmed by analytical ultracentrifugation. These results contribute to a better understanding of this PPI and provide useful insights for drug discovery projects aiming at the modulation of this interaction.

Original languageEnglish
Pages (from-to)1918-1934
Number of pages17
JournalFEBS Journal
Volume288
Issue number6
DOIs
Publication statusPublished - Mar 2021

Funding

The research is supported by funding from the European Union through the TASPPI project (H2020‐MSCA‐ITN‐2015, grant number 675179) and by the LabEx (Laboratory of Excellence) DISTALZ (ANR, ANR‐11‐LABX‐009). The NMR facilities were funded by the Nord Region Council, CNRS, Institut Pasteur de Lille, the European Community (ERDF), the French Ministry of Research and the University of Lille. We acknowledge support for the NMR facilities from TGE RMN THC (CNRS, FR‐3050). We acknowledge Dr. Anders Gunnarsson and MSc Claire Munier, BioPharmaceuticals R&D, AstraZeneca, Gothenburg, Sweden, for fruitful discussions concerning the SPR experiments. We further acknowledge the Professor Christian Ottmann, from the Eindhoven University of Technology, for the creation of the TASPPI consortium and for the continuous suggestions on the progression of the work. We also thank MSc Femke Meijer and the Professor Christian Ottmann, from the Eindhoven University of Technology, for providing us with an aliquot of the diphosphorylated peptide surrounding pS214 + pS324.

FundersFunder number
French Ministry of Research
Nord Region Council
AstraZeneca
Horizon 2020 Framework Programme675179
Université de LilleFR-3050
European CommissionH2020-MSCA-ITN-2015
Eindhoven University of Technology
LabexANR‐11‐LABX‐009
Centre National de la Recherche Scientifique (CNRS)
European Regional Development Fund
Institute Pasteur De Lille

    Keywords

    • 14-3-3 proteins
    • Alzheimer’s disease
    • analytical ultracentrifugation
    • NMR spectroscopy
    • protein–protein interactions
    • Tau protein

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