We present a novel approach for FRET-based detection of protein-protein interactions in the plasma membrane of yeast Saccharomyces cerevisiae using FLIM. FRET (Fluorescence Resonance Energy Transfer) is one of major techniques used to confirm protein-protein interactions within cells.
When FRET between fluorescence donor and acceptor occurs, donor fluorescence lifetime shortens. Monitoring the lifetime shortening of donor fluorescence due to FRET using FLIM (Fluorescence Lifetime IMaging) technique brings a possibility to localize and visualize the protein-protein interaction directly in living cells.
Using these approaches, we show that particular Ato proteins interact with each other in the plasma membrane of cells from colonies occurring in the alkali developmental phase.