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).
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Berridge, M. J. (1993) Inositol trisphosphate and calcium signalling. Nature 361, 315–325.
Downes, P. and Carter, A. N. (1991) Phosphoinositide 3-kinase: a new effector in signal transduction? Cell. Signalling 3, 501–513.
Nahorski, S. R. and Chalhss, R. A. J. (1991) Modulation of receptor-mediated inositol phospholipid breakdown in the brain. Neurochem. Int. 19, 207–212.
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.
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.
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.
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.
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.
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.
Enyedi, P. and Williams, G. H. (1988) Heterogenous inositol tetrakisphosphate binding sites in the adrenal cortex. J. Biol. Chem. 263, 7940–7942.
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.
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.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Humana Press Inc , Totowa, NJ
About this protocol
Cite this protocol
Challiss, R.A.J. (1995). Mass Measurement of Key Phosphoinositide Cycle Intermediates. In: Kendall, D.A., Hill, S.J. (eds) Signal Transduction Protocols. Methods in Molecular Biology™, vol 41. Humana Press. https://doi.org/10.1385/0-89603-298-1:167
Download citation
DOI: https://doi.org/10.1385/0-89603-298-1:167
Publisher Name: Humana Press
Print ISBN: 978-0-89603-298-9
Online ISBN: 978-1-59259-528-0
eBook Packages: Springer Protocols