Affinity methods for phosphorylation-dependent interactions.

Greg Moorhead, Carol MacKintosh

    Research output: Contribution to journalReview articlepeer-review

    4 Citations (Scopus)


    14-3-3s are a highly conserved protein family that exert many regulatory roles in eukaryotic cells by binding to phosphopeptide motifs in diverse target proteins. Here, we describe 14-3-3 affinity binding procedures that can be used to purify and identify 14-3-3-binding phosphoproteins; monitor how their phosphorylation and 14-3-3 binding is regulated by extracellular stimuli; define the functional effects of 14-3-3s on individual targets; and identify relevant protein phosphatases and kinases. In principle, these methods could be adapted to characterize other types of protein-protein interaction that depend on covalent modification of target sites.

    Original languageEnglish
    Pages (from-to)469-478
    Number of pages10
    JournalMethods in Molecular Biology (Clifton, N.J.)
    Publication statusPublished - 1 Jan 2004


    • Protein-protein interactions
    • affinity chromatography
    • phosphorylation
    • phosphopeptide

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics


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