Abstract
Specific membrane proteins, termed lipid flippases, play a central role in facilitating the movement of lipids across cellular membranes. In this protocol, we describe the reconstitution of ATP-driven lipid flippases in liposomes and the analysis of their in vitro flippase activity based on the use of fluorescent lipid derivatives. Working with purified and reconstituted systems provides a well-defined experimental setup and allows to directly characterize these membrane proteins at the molecular level.
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Acknowledgements
This work was supported by the Danish National Research Foundation through the PUMPKIN Center of Excellence (DNRF85), the Danish Council for Independent Research | Natural Sciences (FNU, project number 10-083406), and the Lundbeck Foundation.
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Marek, M., Günther-Pomorski, T. (2016). Assay of Flippase Activity in Proteoliposomes Using Fluorescent Lipid Derivatives. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_18
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DOI: https://doi.org/10.1007/978-1-4939-3179-8_18
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3178-1
Online ISBN: 978-1-4939-3179-8
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