Abstract
Investigations into lipid localization and transport are often hampered by a lack of methods and tools to faithfully visualize lipids in the context of living cells, since fluorescent modifications drastically change lipid properties. Here, we describe the use of bifunctional as well as trifunctional sphingosine to reveal its subcellular localization via crosslinking, fixation, and specific staining by click reaction with a fluorophore. Additionally, these probes allow investigations into lipid metabolism as revealed by thin-layer chromatography.
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References
Hannun YA, Obeid LM (2018) Sphingolipids and their metabolism in physiology and disease. Nat Rev Mol Cell Biol 19:175–191
Quehenberger O, Armando AM, Brown AH et al (2010) Lipidomics reveals a remarkable diversity of lipids in human plasma. J Lipid Res 51:3299–3305
Wang T, Silvius JR (2000) Different sphingolipids show differential partitioning into sphingolipid/cholesterol-rich domains in lipid bilayers. Biophys J 79:1478–1489
Pagano RE, Sleight RG (1985) Defining lipid transport pathways in animal cells. Science 229:1051–1057
Kuerschner L, Ejsing CS, Ekroos K et al (2005) Polyene-lipids: a new tool to image lipids. Nat Methods 2:39–45
Hulce JJ, Cognetta AB, Niphakis MJ et al (2013) Proteome-wide mapping of cholesterol-interacting proteins in mammalian cells. Nat Methods 10:259–264
Haberkant P, Raijmakers R, Wildwater M et al (2013) In vivo profiling and visualization of cellular protein-lipid interactions using bifunctional fatty acids. Angew Chemie—Int Ed 52:4033–4038
Haberkant P, Stein F, Höglinger D et al (2016) Bifunctional sphingosine for cell-based analysis of protein-sphingolipid interactions. ACS Chem Biol 11:222–230
Thiele C, Papan C, Hoelper D et al (2012) Tracing fatty acid metabolism by click chemistry. ACS Chem Biol 7:2004–2011
Höglinger D, Nadler A, Haberkant P et al (2017) Trifunctional lipid probes for comprehensive studies of single lipid species in living cells. Proc Natl Acad Sci U S A 114:1566–1571
Acknowledgments
D.H. is supported by the Deutsche Forschungsgemeinschaft through SFB/TRR 83 and SFB/TRR 186.
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Höglinger, D. (2019). Bi- and Trifunctional Lipids for Visualization of Sphingolipid Dynamics within the Cell. In: Drin, G. (eds) Intracellular Lipid Transport. Methods in Molecular Biology, vol 1949. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9136-5_8
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DOI: https://doi.org/10.1007/978-1-4939-9136-5_8
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-9136-5
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