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Mass Measurement of Key Phosphoinositide Cycle Intermediates

  • R. A John Challiss
Part of the Methods in Molecular Biology™ book series (MIMB, volume 41)

Abstract

The phosphoinositide cycle is now established as a major pathway by which a variety of hormones, neurotransmitters, and other signaling molecules can generate second messengers and thus affect the intracellular milieu (1). Receptor-mediated activation of membrane-associated enzymic activities act on the minor membrane phospholipid phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] to generate three second messenger molecules: inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and 1,2-diacylglycerol via inositol phospholipid-specific phospholipase C (PI-PLC) activity, and phosphatidylinositol 3,4,5-trisphosphate via phosphoinositide 3-kinase activity (2).

Keywords

Wash Buffer Inositol Phosphate Mass Determination Inositol Phospholipid Radioisotopic Label 
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.

References

  1. 1.
    Berridge, M. J. (1993) Inositol trisphosphate and calcium signalling. Nature 361, 315–325.PubMedCrossRefGoogle Scholar
  2. 2.
    Downes, P. and Carter, A. N. (1991) Phosphoinositide 3-kinase: a new effector in signal transduction? Cell. Signalling 3, 501–513.PubMedCrossRefGoogle Scholar
  3. 3.
    Nahorski, S. R. and Chalhss, R. A. J. (1991) Modulation of receptor-mediated inositol phospholipid breakdown in the brain. Neurochem. Int. 19, 207–212.CrossRefGoogle Scholar
  4. 4.
    Challiss, R. A. J., Batty, I. H., and Nahorski, S. R. (1988) Mass measurements of inositol 1,4,5-trisphosphate in rat cerebral cortex slices using a radioreceptor assay. Biochem. Biophys. Res. Commun. 157, 684–691.PubMedCrossRefGoogle Scholar
  5. 5.
    Palmer, S., Hughes, K. T., Lee, D. Y., and Wakelam, M. J. O. (1989) Development of a novel Ins(1,4,5)P3-specific binding assay. Cell. Signalling 1, 147–153.PubMedCrossRefGoogle Scholar
  6. 6.
    Challiss, R. A. J., Chilvers, E. R., Willcocks, A. L., and Nahorski, S. R. (1990) Heterogeneity of [3H]inositol 1,4,5 trisphosphate binding sites in adrenal-cortical membranes. Biochem. J. 265, 421–427.PubMedGoogle Scholar
  7. 7.
    Doné, F. and Reiser, G. (1989) A novel, specific binding protein assay for quantitation of intracellular inositol 1,3,4,5-tetrakisphosphate using a high-affinity receptor from cerebellum. FEBS Lett. 254, 155–158.CrossRefGoogle Scholar
  8. 8.
    Challiss, R. A. J. and Nahorski, S. R. (1990) Neurotransmitter and depolarization-stimulated accumulation of inositol 1,3,4,5-tetrakisphosphate mass in rat cerebral cortex slices. J. Neurochem. 54, 2138–2141.PubMedCrossRefGoogle Scholar
  9. 9.
    Challiss, R. A. J., Willcocks, A. L, Mulloy, B., Potter, B. V. L., and Nahorski, S. R. (1991) Characterization of inositol 1,4,5trisphosphate-and inositol 1,3,4,5-tetra-kisphosphate-binding sites in rat cerebellum. Biochem. J. 274, 861–867.PubMedGoogle Scholar
  10. 10.
    Enyedi, P. and Williams, G. H. (1988) Heterogenous inositol tetrakisphosphate binding sites in the adrenal cortex. J. Biol. Chem. 263, 7940–7942.PubMedGoogle Scholar
  11. 11.
    Sharps, E. S. and McCarl, R. L. (1982) A high-performance liquid chromatographic method to measure 32p-incorporation into phosphorylated metabolites in cultured cells. Anal. Biochem. 124, 421–424.PubMedCrossRefGoogle Scholar
  12. 12.
    Chilvers, E. R., Batty, I. H., Challiss, R. A J., Barnes, P J., and Nahorski, S. R. (1991) Determination of mass changes in phosphatidylinositol 4,5-bisphosphate and evidence for agonist-stimulated metabolism of inositol 1,4,5trisphosphate in airway smooth muscle. Biochem. J. 275, 373–379.PubMedGoogle Scholar
  13. 13.
    Bredt, D. S., Mourey, R. J., and Snyder, S. H. (1989) A simple, sensitive and specific assay for inositol 1,4,Strisphosphate in biological tissues. Biochem. Biophys. Res. Commun. 159, 976–982.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc , Totowa, NJ 1995

Authors and Affiliations

  • R. A John Challiss
    • 1
  1. 1.Department of Cell Physiology and PharmacologyUniversity of LeicesterUK

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