Understanding GPCR Signaling Using TRUPATH
Type
Blog Post
...was developed by constructing 14 luciferase donor Rluc8-Gα chimeras, four acceptor GFP2-Gγ chimeras, and...protein. For the Gα constructs, localization of the Rluc8 was more challenging. The Roth lab used structure-guided...refinement to determine the optimal localization of the Rluc8 insertion within the Gα. Despite exhaustive refinement...refinement, it was not possible to generate an optimal Rluc8-Gα construct for two of the 16 non-visual G protein... plasmids expressing the GPCR of interest, a Gα-Rluc8 plasmid, and the appropriate Gγ-GFP2 and Gβ constructs...inactive state, the luminescent chimeric protein Gα-Rluc8 and Gγ-GFP2 will be in close association, resulting...BRET2 ratio, which is the ratio of GFP2 emission to RLuc8 emission (see figure).
What can TRUPATH be used ...