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

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Etat: Public
Version: de l'auteur⸱e
Licence: Non spécifiée
ID Serval
serval:BIB_A9847654B936
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles.
Périodique
Methods in molecular biology
Auteur⸱e⸱s
D'Agostino M., Mayer A.
ISSN
1940-6029 (Electronic)
ISSN-L
1064-3745
Statut éditorial
Publié
Date de publication
2019
Peer-reviewed
Oui
Volume
1860
Pages
253-262
Langue
anglais
Notes
Publication types: Journal Article
Publication Status: ppublish
Résumé
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.
Mots-clé
Lipid mixing, Membrane fusion, Vacuole, Yeast
Pubmed
Financement(s)
Fonds national suisse / Projets / 31003A_179306
Création de la notice
05/11/2018 9:15
Dernière modification de la notice
15/01/2021 7:26
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