A live cell assay of GPCR coupling allows identification of optogenetic tools for controlling Go and Gi signaling.

ABSTRACT:

Background

Animal opsins are light-sensitive G-protein-coupled receptors (GPCRs) that enable optogenetic control over the major heterotrimeric G-protein signaling pathways in animal cells. As such, opsins have potential applications in both biomedical research and therapy. Selecting the opsin with the best balance of activity and selectivity for a given application requires knowing their ability to couple to a full range of relevant Gα subunits. We present the GsX assay, a set of tools based on chimeric Gs subunits that transduce coupling of opsins to diverse G proteins into increases in cAMP levels,  measured with a real-time reporter in living cells. We use this assay to compare coupling to Gi/o/t across a panel of natural and chimeric opsins selected for potential application in

SUBMITTER: Ballister ER 

PROVIDER: S-EPMC5771134 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature