Abstract
Acidic phospholipids are minor membrane lipids but critically important for signaling events. The main acidic phospholipids are phosphatidylinositol phosphates (PIPs also known as phosphoinositides), phosphatidylserine (PS), and phosphatidic acid (PA). Acidic phospholipids are precursors of second messengers of key signaling cascades or are second messengers themselves. They regulate the localization and activation of many proteins, and are involved in virtually all membrane trafficking events. As such, it is crucial to understand the subcellular localization and dynamics of each of these lipids within the cell. Over the years, several techniques have emerged in either fixed or live cells to analyze the subcellular localization and dynamics of acidic phospholipids. In this chapter, we review one of them: the use of genetically encoded biosensors that are based on the expression of specific lipid binding domains (LBDs) fused to fluorescent proteins. We discuss how to design such sensors, including the criteria for selecting the lipid binding domains of interest and to validate them. We also emphasize the care that must be taken during data analysis as well as the main limitations and advantages of this approach.
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Acknowledgement
We thank Mathilde Simon, Marie-Cécile Caillaud, and Marlene Dreux for commenting the manuscript. Y.J. has received funding from the European Research Council—ERC Grant Agreement no. [3363360-APPL] and from the Marie Curie Action—CIG Grant Agreement no. [PCIG-GA-2011-303601] under the European Union’s Seventh Framework Programme (FP/2007-2013).
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Platre, M.P., Jaillais, Y. (2016). Guidelines for the Use of Protein Domains in Acidic Phospholipid Imaging. In: Waugh, M. (eds) Lipid Signaling Protocols. Methods in Molecular Biology, vol 1376. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3170-5_15
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