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. 2015 Mar 19;50(3):231–241. doi: 10.3109/10409238.2015.1023252

Figure 1.

Figure 1.

Ensemble vesicle fusion assays. (A) Vesicle–vesicle fusion assay using fluorescently labeled lipid analogs (Struck et al., 1981). Vesicles containing a high concentration of NBD-PE and Rh-PE are characterized by quenching of NBD fluorescence due to FRET between NBD and rhodamine. Upon fusion to unlabeled vesicles the concentration of NBD and rhodamine decreases, resulting in less FRET efficiency and thus a consequent increase in NBD fluorescence (dequenching). (B) Application of the ensemble lipid-mixing vesicle fusion assay to proteoliposomes containing SNARE proteins (Weber et al., 1998). NBD-PE and Rh-PE containing vesicles reconstituted with v-SNARES (synaptobrevin shown in blue) fuse with unlabeled vesicle containing t-SNAREs (syntaxin shown in red and SNAP-25 shown in green), resulting in the dequenching of NBD fluorescence.