ArticleMethods in molecular biology (Clifton, N.J.)2025
Measuring Phospholipid Flippase Activity by NBD-Lipid Uptake in Living Yeast Cells.
Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Phospholipid flippases in the P4-ATPase family are essential for establishing membrane asymmetry. These ATP-powered pumps translocate specific lipids from the exofacial leaflet to the cytosolic leaflet of the plasma membrane, thereby concentrating substrate lipids, such as phosphatidylserine, in the cytosolic leaflet while non-substrate lipids populate the exofacial leaflet. Here, we describe a method for measuring P4-ATPase transport activity in the yeast plasma membrane by using flow cytometry to quantify the uptake of lipids derivatized with a fluorescent [7-nitro-2-1,3-benzoxadiazol-4-yl)amino] (NBD) group on a short (C6) fatty acyl chain. The NBD-lipid uptake assay quantitatively measures P4-ATPase transport activity and substrate selectivity in the native membrane environment.
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