| A Cell-Based Drug Screening Strategy For G Protein Coupled Receptors |
|
Presentation: 3B03 Session: Target Biology & Screening: GPCRsBrian F. O'Dowd, University of Toronto Presenting Author: Brian O'Dowd, University of Toronto - Canada We describe the Multipurpose Original Assay (MOCA), a drug screening strategy suitable for the discovery of compounds targeting G protein coupled receptors (GPCRs). Incorporation of a nuclear localization sequence (NLS) in a critical location resulted in GPCR translocation from the cell surface to the nucleus in a ligand-independent and time-dependent manner. Antagonist blockade of the GPCR-NLS blocked nuclear translocation in a sensitive dose-dependent manner, and thus this MOCA method can be utilized for identification of compounds targeting orphan GPCRs. Nuclear trafficking of the GPCR-NLS was visualized by fusion to optically detectable molecules, or residual cell surface receptor densities measured by radiolabeled ligand or fluorescence detection in a multiwell format, suitable for high throughput screening. Thus, we describe a novel cellular MOCA method for the effective screening of antagonist drugs targeting GPCRs. |