원문정보
초록
영어
Taste receptors belong to the subfamily of G-protein-coupled receptors (GPCR). When the GPCR binds to the ligand, recruitment of β-arrestin (β-arr) to agonist-stimulated receptor occurs. In this study, HEK-293 cells which coexpress the taste receptor T2R38 and β-arr2 was established,
and we observed that the recruitment of β-arr2 to the C-terminus of receptor was stimulated by the specific ligand. The monitoring method was based on the fluorescence resonance energy transfer (FRET) from GFP to DsRed, which were attached to the c-terminals of β-arr2 and
T2R38, respectively. The fluorescence of GFP and DsRed excited at 485nm was monitored at 535nm and 590nm, respectively. Agonist-induced signals were measured by the DsRed/GFP emission ratio. Mammalian cell-derived nanovesicles containing taste receptor and β-arr2 were
formed by treating the HEK-293 cells with cytochalasin B. The taste receptor and β-arr2 imbedded in the nanovesicles were confirmed by the fluorescence image using confocal microscopy. This methodology would be very effective for the development of GPCR-based biosensor, which can be used for ligand screening.