SYP42 and SYP43 can drive liposome fusion. Purified SYP41 (A), SYP42 (B), or SYP43 (C) was incorporated into acceptor vesicles (non-fluorescent vesicles), and VTI12 was reconstituted into donor vesicles containing fluorescent lipids. Membrane fusion with (black) or without (red) YKT62 was measured by monitoring NBD fluorescence with excitation wavelength of 460 nm and emission wavelength of 530 nm. The fluorescence was recorded every ten seconds for 10 min. The maximal fluorescence intensity (MFI) was obtained by adding 1 μl of 10% (v/v) Triton X-100, and recorded fluorescence measurements were normalized to the MFI. D. Schematic diagram of the assays shown in Figures 3 and 4.
Kim and Bassham BMC Biochemistry 2013 14:22 doi:10.1186/1471-2091-14-22