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Assay of Flippase Activity in Proteoliposomes Using Fluorescent Lipid Derivatives

  • Protocol
P-Type ATPases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1377))

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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Correspondence to Thomas Günther-Pomorski .

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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

  • eBook Packages: Springer Protocols

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