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Analysis of Cellular Phosphoinositides and Phosphoinositols by Extraction and Simple Analytical Procedures

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Biomembrane Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 27))

Abstract

Part A. Biosynthesis and Extraction of Phosphoinositides and Phosphoinositols

The minor inositol-containing membrane phospholipids, the phosphoinositides, play a central role in cell signal transduction. Activation of a hormone-sensitive phospholipase C (phosphoinositidase C) results in the rapid catabolism of the polyphosphoinositides to form the two second messengers inositol 1,4,5-trisphosphate (Ins(l,4,5)P3), a water soluble phosphoinositol that promotes the release of Ca2+ from intracellular stores, and diacylglycerol (DG), which remains in the plasma membrane and activates protein kinase C (1–3). See Fig. 1 for a summary of the pathways.

Metabolic pathways activated as a consequence of phosphoinositidase C action. In Panel A major metabolic pathways activated by phosphoinositidase C action on PtdIns(4,5)P2 are shown with solid arrows. Some of the additional pathways that may be activated are shown by broken arrows Abbreviations: Ptdlns: phosphatidylinositol; PtdIns4P. phosphatidylinositol 4-phosphate; PtdIns(4,5)P2 phosphatidylinositol 4,5-bisphosphate; DG: diacylglycerol; PtdOH: phosphatidic acid; CDP-DG: CDP-diacylglycerol; Ins: Inositol. For phosphoinositols, abbreviations are in the form Ins(x,y,z)Pn where x, y, and z refer to the positions of the phosphate groups on the myo-inositol ring and n refers to the total number of phosphates. Panel B is a simplified outline of the metabolic pathways in (A) to show alternative abbreviations. PI: phosphatidylinositol, PIP: phosphatidyl-inositol phosphate; PIP2, phosphatidylinositol bisphosphate; DG, PtdOH, CDP-DG and Ins as above. Phosphoinositols are referred to as IPn where n refers to the number of phosphates on the inositol ring In both panels, sites of Li*+ inhibition are also shown.

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Authors and Affiliations

  1. University Department of Clinical Chemistry, The Royal Infirmary, Edinburgh, Scotland (Current address: Cecil H. and Ida Green Center, University of Texas Southwestern Medical Center, Dallas, TX

    Ian M. Bird

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© 1994 Humana Press Inc. Totowa, NJ

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Bird, I.M. (1994). Analysis of Cellular Phosphoinositides and Phosphoinositols by Extraction and Simple Analytical Procedures. In: Biomembrane Protocols. Methods in Molecular Biology, vol 27. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-250-7:227

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  • DOI: https://doi.org/10.1385/0-89603-250-7:227

  • Publisher Name: Springer, Totowa, NJ

  • Print ISBN: 978-0-89603-250-7

  • Online ISBN: 978-1-59259-514-3

  • eBook Packages: Springer Protocols

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