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)

Research output: Contribution to journalArticleAcademicpeer-review

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
Number of pages17
JournalFEBS Journal
DOIs
Publication statusAccepted/In press - 26 Sep 2020

Keywords

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

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