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2019
- doi: 10.1007/978-1-4939-8760-3_16
- issn: 1064-3745
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info:eu-repo/semantics/altIdentifier/doi/10.1007/978-1-4939-8760-3_16
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info:eu-repo/semantics/altIdentifier/pmid/30317510
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info:eu-repo/semantics/altIdentifier/eissn/1940-6029
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info:eu-repo/grantAgreement/SNF/Projects/31003A_179306///
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info:eu-repo/semantics/altIdentifier/urn/urn:nbn:ch:serval-BIB_A9847654B9368
info:eu-repo/semantics/openAccess , CC BY 4.0 , https://creativecommons.org/licenses/by/4.0/
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D'Agostino M. et al., « Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles. », Serveur académique Lausannois, ID : 10.1007/978-1-4939-8760-3_16
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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.