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Measurement of Phospholipase C by Monitoring Inositol Phosphates Using [3H]Inositol-Labeling Protocols in Permeabilized Cells
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.
Affiliation(s): (2) Department of Physiology, University College London, London, UK
Series: Methods in Molecular Biology  |  Volume: 312  |  Pub. Date: Aug-15-2005  |  Page Range: 183-193  |  DOI: 10.1385/1-59259-949-4:183
Subject:  Cell Biology
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