Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles.

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State: Public
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Serval ID
serval:BIB_A9847654B936
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles.
Journal
Methods in molecular biology
Author(s)
D'Agostino M., Mayer A.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Publication state
Published
Issued date
2019
Peer-reviewed
Oui
Volume
1860
Pages
253-262
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Fluorescence de-quenching can be used to analyze membrane lipid mixing during an in vitro fusion reaction. Here we describe a method to measure lipid mixing using vacuolar membranes purified from the yeast Saccharomyces cerevisiae. Labeling the isolated organelles with rhodamine-phosphatidylethanolamine allows to reveal ATP-dependent lipid mixing through fluorescence de-quenching in a spectrofluorometer. Combining this assay with content mixing indicators, such as the fusion-dependent maturation of a luminal vacuolar phosphatase, then permits the detection of hemifusion intermediates and the analysis of the requirements for fusion pore opening.
Keywords
Lipid mixing, Membrane fusion, Vacuole, Yeast
Pubmed
Funding(s)
Swiss National Science Foundation / Projects / 31003A_179306
Create date
05/11/2018 8:15
Last modification date
15/01/2021 6:26
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