Recognition of an intra-chain tandem 14-3-3 binding site within PKC epsilon

Brenda Kostelecky, Adrian T. Saurin, Andrew Purkiss, Peter J. Parker, Neil Q. McDonald (Lead / Corresponding author)

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    Abstract

    The phosphoserine/threonine binding protein 14-3-3 stimulates the catalytic activity of protein kinase C-epsilon (PKC epsilon) by engaging two tandem phosphoserine-containing motifs located between the PKC epsilon regulatory and catalytic domains (V3 region). Interaction between 14-3-3 and this region of PKC epsilon is essential for the completion of cytokinesis. Here, we report the crystal structure of 14-3-3 zeta bound to a synthetic diphosphorylated PKC epsilon V3 region revealing how a consensus 14-3-3 site and a divergent 14-3-3 site cooperate to bind to 14-3-3 and so activate PKC epsilon. Thermodynamic data show a markedly enhanced binding affinity for two-site phosphopeptides over single-site 14-3-3 binding motifs and identifies Ser 368 as a gatekeeper phosphorylation site in this physiologically relevant 14-3-3 ligand. This dual-site intra-chain recognition has implications for other 14-3-3 targets, which seem to have only a single 14-3-3 motif, as other lower affinity and cryptic 14-3-3 gatekeeper sites might exist.

    Original languageEnglish
    Pages (from-to)983-989
    Number of pages7
    JournalEMBO Reports
    Volume10
    Issue number9
    DOIs
    Publication statusPublished - Sep 2009

    Keywords

    • PROTEIN
    • MEMBRANE H+-ATPASE
    • crystal structure
    • CRYSTAL-STRUCTURE
    • DIFFRACTION DATA
    • SOFTWARE
    • ACTIVATION
    • PEPTIDE
    • PKC epsilon
    • REQUIRES PHOSPHORYLATION
    • 14-3-3
    • ASSOCIATION
    • INHIBITION
    • isothermal titration calorimetry

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