Abstract
Several mechanisms deliver specific lipid types from one organelle to another in cells. Two distinct mechanisms exist for inter-organelle lipid trafficking in eukaryotic cells: vesicular and non-vesicular. Lipid-transfer proteins (LTPs) that catalyze the inter-membrane transfer of lipids play pivotal roles in non-vesicular systems. As a key biochemical feature, LTPs extract a specific lipid type from membranes in vitro. This chapter describes two assay systems to assess the lipid extraction activity of the ceramide transport protein CERT from membranes under cell-free conditions: one is a conventional assay system using radioactive lipid ligands, while the other is a more recently developed assay system using surface plasmon resonance. These methods are applicable to other LTPs.
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Acknowledgments
We deeply thank our coworkers for their invaluable contributions to studies concerning CERT. This work was supported by Takeda Science Foundation, and AMED-CREST (to K.H.), and the Ministry of Economy, Trade, and Industry (METI) and the New Energy and Industrial Technology Development Organization (NEDO) of Japan (to S.T.).
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Hanada, K., Sugiki, T. (2017). In Vitro Assay to Extract Specific Lipid Types from Phospholipid Membranes Using Lipid-Transfer Proteins: A Lesson from the Ceramide Transport Protein CERT. In: Wood, P. (eds) Lipidomics. Neuromethods, vol 125. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6946-3_6
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DOI: https://doi.org/10.1007/978-1-4939-6946-3_6
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