biostudies-literatureUnknownFranchini LNIDCD NIH HHSNHLBI NIH HHSG protein-coupled receptors (GPCRs) are key pharmacological targets, yet many remain underutilized due to unknown activation mechanisms and ligands. Orphan GPCRs, lacking identified natural ligands, are a high priority for research, as identifying their ligands will aid in understanding their functions and potential as drug targets. Most GPCRs, including orphans, couple to G_i/o/z family members, however current assays to detect their activation are limited, hindering ligand identification efforts. We introduce G_ZESTY, a highly sensitive, cell-based assay developed in an easily deliverable format designed to study the pharmacology of G_i/o/z-coupled GPCRs and assist in deorphanization. We optimized assay conditions and developed an all-in-one vector employingbioRxiv : the preprint server for biology2024.07.26.605282https://www.ebi.ac.uk/biostudies/studies/S-EPMC11291178biostudies-literatureGz Enhanced Signal Transduction assaY (G_ZESTY) for GPCR deorphanization.PMC11291178R01 DC022104R01 HL153988Porter JJLueck JDOrlandi CFranchini LfalseGz Enhanced Signal Transduction assaY (G_ZESTY) for GPCR deorphanization.G protein-coupled receptors (GPCRs) are key pharmacological targets, yet many remain underutilized due to unknown activation mechanisms and ligands. Orphan GPCRs, lacking identified natural ligands, are a high priority for research, as identifying their ligands will aid in understanding their functions and potential as drug targets. Most GPCRs, including orphans, couple to G_i/o/z family members, however current assays to detect their activation are limited, hindering ligand identification efforts. We introduce G_ZESTY, a highly sensitive, cell-based assay developed in an easily deliverable format designed to study the pharmacology of G_i/o/z-coupled GPCRs and assist in deorphanization. We optimized assay conditions and developed an all-in-one vector employing2024-01-01T00:00:00Z2024 Jul2026-06-24T03:22:00.86Z2024-10-17T23:03:34.29ZS-EPMC112911783909186910.1101/2024.07.26.605282