Cell-based lipid flippase assay employing fluorescent lipid derivatives

Maria Stumph Jensen, Sara Costa, Thomas Günther-Pomorski, Rosa Laura Lopez Marques

2 Citations (Scopus)

Abstract

P-type ATPases in the P4 subfamily (P4-ATPases) are transmembrane proteins unique for eukaryotes that act as lipid flippases, i.e., to translocate phospholipids from the exofacial to the cytofacial monolayer of cellular membranes. While initially characterized as aminophospholipid translocases, studies of individual P4-ATPase family members from fungi, plants, and animals show that P4-ATPases differ in their substrate specificities and mediate transport of a broader range of lipid substrates. Here, we describe an assay based on fluorescent lipid derivatives to monitor and characterize lipid flippase activities in the plasma membrane of cells, using yeast as an example.

Original languageEnglish
Title of host publicationP-Type ATPases : methods and protocols
EditorsMaike Bublitz
Number of pages12
VolumeVI
PublisherSpringer
Publication date1 Jan 2016
Pages371-382
Chapter33
ISBN (Print)978-1-4939-3178-1
ISBN (Electronic)978-1-4939-3179-8
DOIs
Publication statusPublished - 1 Jan 2016
SeriesMethods in Molecular Biology
Volume1377
ISSN1064-3745

Keywords

  • Azoles
  • Enzyme Assays
  • Flow Cytometry
  • Fluorescent Dyes
  • Lipid Metabolism
  • Lipids
  • Nitrobenzenes
  • Phospholipid Transfer Proteins
  • Saccharomyces cerevisiae
  • Journal Article

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