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Membrane fusion mediated by peptidic SNARE protein analogues: Evaluation of FRET‐based bulk leaflet mixing assays
Author(s) -
Hubrich Barbara E.,
Wehland JanDirk,
Groth Mike C.,
Schirmacher Anastasiya,
Hubrich Raphael,
Steinem Claudia,
Diederichsen Ulf
Publication year - 2021
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.3327
Subject(s) - förster resonance energy transfer , liposome , lipid bilayer fusion , biophysics , chemistry , membrane , fusion , transmembrane protein , peptide , lipid bilayer , fusion protein , biochemistry , receptor , biology , fluorescence , recombinant dna , physics , linguistics , philosophy , quantum mechanics , gene
Peptide‐mediated membrane fusion is frequently studied with in vitro bulk leaflet mixing assays based on Förster resonance energy transfer (FRET). In these, customized liposomes with fusogenic peptides are equipped with lipids which are labeled with fluorophores that form a FRET pair. Since FRET is dependent on distance and membrane fusion comes along with lipid mixing, the assays allow for conclusions on the membrane fusion process. The experimental outcome of these assays, however, greatly depends on the applied parameters. In the present study, the influence of the peptides, the size of liposomes, their lipid composition and the liposome stoichiometry on the fusogenicity of liposomes are evaluated. As fusogenic peptides, soluble N ‐ethylmaleimide‐sensitive‐factor attachment receptor (SNARE) protein analogues featuring artificial recognition units attached to the native SNARE transmembrane domains are used. The work shows that it is important to control these parameters in order to be able to properly investigate the fusion process and to prevent undesired effects of aggregation.