Effects of lipid kinase expression and cellular stimuli on phosphatidylinositol 5-phosphate levels in mammalian cell lines

Hilary F. Roberts, Jonathan H. Clarke, Andrew J. Letcher, Robin F. Irvine, Katherine A. Hinchliffe

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Phosphatidylinositol 5-phosphate (PtdIns5P) is a relatively recently discovered inositol lipid whose metabolism and functions are not yet clearly understood. We have transfected cells with a number of enzymes that are potentially implicated in the synthesis or metabolism of PtdIns5P, or subjected cells to a variety of stimuli, and then measured cellular PtdIns5P levels by a specific mass assay. Stable or transient overexpression of Type IIα PtdInsP kinase, or transient overexpression of Type Iα or IIβ PtdInsP kinases caused no significant change in cellular PtdIns5P levels. Similarly, subjecting cells to oxidative stress or EGF stimulation had no significant effect on PtdIns5P, but stimulation of HeLa cells with a phosphoinositide- specific PLC-coupled agonist, histamine, caused a 40% decrease within 1 min. Our data question the degree to which inositide kinases regulate PtdIns5P levels in cells, and we discuss the possibility that a significant part of both the synthesis and removal of this lipid may be regulated by phosphatases and possibly phospholipases. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
    Original languageEnglish
    Pages (from-to)2868-2872
    Number of pages4
    JournalFEBS Letters
    Volume579
    Issue number13
    DOIs
    Publication statusPublished - 23 May 2005

    Keywords

    • Mass assay
    • Phosphatidylinositol
    • Phosphatidylinositol 5-phosphate
    • Type II phosphatidylinositol phosphate kinase

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