Author:
Mathiassen Patricia P. M.,Menon Anant K.,Pomorski Thomas Günther
Abstract
AbstractTransbilayer movement of phospholipids in biological membranes is mediated by a diverse set of lipid transporters. Among them are scramblases that facilitate a rapid bi-directional movement of lipids without metabolic energy input. Here, we established a new fluorescence microscopy-based assay for detecting phospholipid scramblase activity of membrane proteins upon their reconstitution into giant unilamellar vesicles formed from proteoliposomes by electroformation. The assay is based on chemical bleaching of fluorescence of a photostable ATTO-dye labeled phospholipid with the membrane-impermeant reductant sodium dithionite. We demonstrate that this new methodology is suitable for the study of the scramblase activity of the yeast endoplasmic reticulum at single vesicle level.
Funder
German research foundation
German Research Foundation
DAAD
Ruhr-Universität Bochum
Publisher
Springer Science and Business Media LLC
Cited by
5 articles.
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