Structural insights into the regulation of PDK1 by phosphoinositides and inositol phosphates

David Komander, Alison Fairservice, Maria Deak, Gursant S. Kular, Alan R. Prescott, C. Peter Downes, Stephen T. Safrany, Dario R. Alessi, Daan M.F. Van Aalten

    Research output: Contribution to journalArticlepeer-review

    150 Citations (Scopus)


    3-phosphoinositide-dependent protein kinase-1 (PDK1) phosphorylates and activates many kinases belonging to the AGC subfamily. PDK1 possesses a C-terminal pleckstrin homology (PH) domain that interacts with PtdIns(3,4,5)P3/PtdIns(3,4)P2 and with lower affinity to PtdIns(4,5)P2. We describe the crystal structure of the PDK1 PH domain, in the absence and presence of PtdIns(3,4,5)P3 and Ins(1,3,4,5)P4. The structures reveal a 'budded' PH domain fold, possessing an N-terminal extension forming an integral part of the overall fold, and display an unusually spacious ligand-binding site. Mutagenesis and lipid-binding studies were used to define the contribution of residues involved in phosphoinositide binding. Using a novel quantitative binding assay, we found that Ins(1,3,4,5,6)P5 and InsP6, which are present at micromolar levels in the cytosol, interact with full-length PDK1 with nanomolar affinities. Utilising the isolated PDK1 PH domain, which has reduced affinity for Ins(1,3,4,5,6)P5/InsP6, we perform localisation studies that suggest that these inositol phosphates serve to anchor a portion of cellular PDK1 in the cytosol, where it could activate its substrates such as p70 S6-kinase and p90 ribosomal S6 kinase that do not interact with phosphoinositides.

    Original languageEnglish
    Pages (from-to)3918-3928
    Number of pages11
    JournalEMBO Journal
    Issue number20
    Publication statusPublished - 13 Oct 2004


    • Inositol phosphates
    • Phosphoinositides
    • PI-3 kinase pathway
    • Protein crystallography
    • Protein structure

    ASJC Scopus subject areas

    • General Neuroscience
    • Molecular Biology
    • General Immunology and Microbiology
    • General Biochemistry,Genetics and Molecular Biology


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