A New Chemical Probe for Phosphatidylinositol Kinase Activity

Authors

  • Dr. Allison R. Sherratt,

    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    Search for more papers by this author
  • Neda Nasheri,

    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    2. Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5 (Canada)
    Search for more papers by this author
  • Dr. Craig S. McKay,

    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    Search for more papers by this author
  • Shifawn O'Hara,

    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    Search for more papers by this author
  • Ashley Hunt,

    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    Search for more papers by this author
  • Dr. Zhibin Ning,

    1. Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5 (Canada)
    Search for more papers by this author
  • Prof. Daniel Figeys,

    1. Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5 (Canada)
    Search for more papers by this author
  • Prof. Natalie K. Goto,

    1. Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5 (Canada)
    Search for more papers by this author
  • Prof. John Paul Pezacki

    Corresponding author
    1. Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)
    2. Department of Biochemistry, Microbiology and Immunology, Ottawa Institute of Systems Biology, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5 (Canada)
    • Life Sciences Division, National Research Council of Canada, 100 Sussex Drive, Ottawa, K1A 0R6 (Canada)===

    Search for more papers by this author

Abstract

Phosphatidylinositol kinases (PIKs) are key enzymatic regulators of membrane phospholipids and membrane environments that control many aspects of cellular function, from signal transduction to secretion, through the Golgi apparatus. Here, we have developed a photoreactive “clickable” probe, PIK-BPyne, to report the activity of PIKs. We investigated the selectivity and efficiency of the probe to both inhibit and label PIKs, and we compared PIK-BPyne to a wortmannin activity-based probe also known to target PIKs. We found that PIK-BPyne can act as an effective in situ activity-based probe, and for the first time, report changes in PI4K-IIIβ activity induced by the hepatitis C virus. These results establish the utility of PIK-BPyne for activity-based protein profiling studies of PIK function in native biological systems.

Ancillary