| 摘要 |
Use of mutated beta-arrestin for an improved enzyme complementation assay or translocation assay, the improved enzyme complementation assay comprising: i) adding a substrate to a cell comprising a GPCR-EA fusion protein and a beta-arrestin-EB fusion protein, wherein the beta-arrestin is mutated, ii) adding a ligand to obtain, if possible, a GPCR-EA/beta-arrestin-EB complex, and iii) measuring a signal arising from association of EA and EB to create an enzymatically active protein catalyzing conversion of the substrate which leads to a detectable signal, wherein the improvement leads to an increased signal compared with the signal obtained by use of the same process employing a beta-arrestin-EB fusion protein, wherein the beta-arrestin is wild type beta-arrestin, and the improved beta-arrestin translocation assay comprising i) providing a cell expressing a GPCR and comprising a beta-arrestin associated with an optically detectable molecule, ODM, wherein the beta-arrestin is mutated, ii) adding a ligand to obtain, if possible, a GPCR/beta-arrestin complex, and iii) detecting a translocation of the optically detectable molecule, wherein the improvement leads to a increased and prolonged translocation of the beta-arrestin associated with an optically detectable molecule as compared with the signal obtained by use of the same assay employing a beta-arrestin associated with an optically detectable molecule, wherein the beta-arrestin is wild type beta-arrestin. |
