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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)

Abstract

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.

Keywords

HL60 Cell Inositol Phosphate Ethylene Glycol Tetraacetic Acid Ethylene Glycol Tetraacetic Acid Assay Tube 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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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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