Assay of Flippase Activity in Proteoliposomes Using Fluorescent Lipid Derivatives.

Magdalena Marek, Thomas Günther-Pomorski

Journal: Methods in molecular biology (Clifton, N.J.) 2016;1377():181-91

PMID: 26695033

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.

Address: Department of Plant and Environmental Sciences, Centre for Membrane Pumps in Cells and Disease (PUMPKIN), University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark.; Department of Plant and Environmental Sciences, Centre for Membrane Pumps in Cells and Disease (PUMPKIN), University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Denmark. [email protected].

DOI: 10.1007/978-1-4939-3179-8_18

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MeSH Terms: ATP-Binding Cassette Transporters, Adenosine Triphosphate, Biological Transport, Cell Membrane, Dithionite, Fluorescence, Lipids, Membrane Proteins, Molecular Biology, Proteolipids