Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol-Labeling Protocols in Permeabilized Cells

  • Alison Skippen
  • Philip Swigart
  • Shamshad Cockcroft
Part of the Methods in Molecular Biology™ book series (MIMB, volume 312)


Hormones, neurotransmitters, chemoattractants, and growth factors all elicit intracellular responses on binding to cell surface receptors by activating inositol phospholipid-specific phospholipase C (PLC). Activated PLC catalyzes the hydrolysis of phosphatidylinositol bisphosphate (PIP2), a minor membrane phospholipid, to form two second messengers, diacylglycerol (DAG) and inositol (1,4,5)trisphosphate [Ins(1,4,5,)P3]. DAG is a direct activator of protein kinase C isozymes, and Ins(1,4,5)P3 mobilizes intracellular Ca2+. G protein-coupled receptors couple to the PLC-β family via G proteins, and tyrosine kinase receptors activate PLC-γ isozymes (1,2). Regardless of the PLC isozyme activated, the product is invariantly Ins(1,4,5)P3.


HL60 Cell Inositol Phosphate Ethylene Glycol Tetraacetic Acid Ethylene Glycol Tetraacetic Acid Assay Tube 
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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Alison Skippen
    • 1
  • Philip Swigart
    • 1
  • Shamshad Cockcroft
    • 1
  1. 1.Department of PhysiologyUniversity College LondonLondon

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