Abstract
Lipid phosphate molecules such as phosphatidate, lysophosphatidate, and diacylglycerol pyrophosphate play roles as signaling molecules in prokaryotic and eukaryotic cells. The cellular processes by which lipid phosphate molecules signal may be attenuated through the action of lipid phosphate phosphatase enzymes. The levels of lipid phosphate phosphatase activities may be used as a marker of signaling events in the cell. In this chapter we describe enzymatic assays that are routinely used to measure the activities of phosphatidate phosphatase, lysophosphatidate phosphatase, and diacylglycerol pyrophosphate phosphatase. These activities are measured by following the release of water-soluble radioactive inorganic phosphate from chloroform-soluble radioactive lipid phosphate substrate following a simple chloroform/methanol/water phase partition.
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References
Brindley, D. N. and Waggoner, D. W. (1998) Mammalian lipid phosphate phosphohydrolases. J. Biol. Chem. 273, 24,281–24,284.
Carman, G. M. (1997) Phosphatidate phosphatases and diacylglycerol pyrophosphate phosphatases in Saccharomyces cerevisiae and Escherichia coli. Biochim. Biophys. Acta 1348, 45–55.
Kanoh, H., Kai, M., and Wada, I. (1997) Phosphatidic acid phosphatase from mammalian tissues: discovery of channel-like proteins with unexpected functions. Biochim. Biophys. Acta 1348, 56–62.
Stukey, J. and Carman, G. M. (1997) Identification of a novel phosphatase sequence motif. Protein Sci. 6, 469–472.
Neuwald, A. F. (1997) An unexpected structural relationship between integral membrane phosphatases and soluble haloperoxidases. Protein Sci. 6, 1764–1767.
Hemrika, W., Renirie, R., Dekker, H. L., et al. (1997) From phosphatases to vanadium peroxidases: a similar architecture of the active site. Proc. Nat. Acad. Sci. USA 94, 2145–2149.
Jamal, Z., Martin, A., Gomez-Munoz, A., and Brindley, D. N. (1991) Plasma membrane fractions from rat liver contain a phosphatidate phosphohydrolase distinct from that in the endoplasmic reticulum and cytosol. J. Biol. Chem. 266, 2988–2996.
Brindley, D. N. (1984) Intracellular translocation of phosphatidate phosphohydrolase and its possible role in the control of glycerolipid synthesis. Prog. Lipid Res. 23, 115–133.
Brindley, D. N., English, D., Pilquil, C., et al. (2002) Lipid phosphate phosphatases regulate signal transduction through glycerolipids and sphingolipids. Biochim. Biophys. Acta. 1582, 33–44.
Sciorra, V. A. and Morris, A. J. (2002) Roles for lipid phosphate phosphatases in regulation of cellular signaling. Biochim. Biophys. Acta 1582, 45–51.
Walsh, J. P. and Bell, R. M. (1986) sn-1,2-diacylglycerol kinase of Escherichia coli. Structural and kinetic analysis of the lipid cofactor dependence. J. Biol. Chem. 261, 15,062–15,069.
Wissing, J. B. and Behrbohm, H. (1993) Phosphatidate kinase, a novel enzyme in phospholipid metabolism. Purification, subcellular localization, and occurrence in the plant kingdom. Plant Physiol. 102, 1243–1249.
Furneisen, J. M. and Carman, G. M. (2000) Enzymological properties of the LPP1-encoded lipid phosphatase from Saccharomyces cerevisiae. Biochim. Biophys. Acta 1484, 71–82.
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© 2004 Humana Press Inc., Totowa, NJ
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Han, GS., Carman, G.M. (2004). Assaying Lipid Phosphate Phosphatase Activities. In: Dickson, R.C., Mendenhall, M.D. (eds) Signal Transduction Protocols. Methods in Molecular Biology, vol 284. Humana Press. https://doi.org/10.1385/1-59259-816-1:209
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DOI: https://doi.org/10.1385/1-59259-816-1:209
Publisher Name: Humana Press
Print ISBN: 978-1-58829-245-2
Online ISBN: 978-1-59259-816-8
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