Antagonists Enhance Cell-Surface Expression of Mammalian Odorant Receptors

Functional characterization of vertebrate odorant receptors (ORs), members of the G protein-coupled receptor (GPCR) family, is essential for understanding olfaction. However, the functional expression of ORs in heterologous cells is often challenging, at least partly caused by structural instability in non-olfactory cells. Antagonists have been shown to restore membrane expression of some non-olfactory GPCR mutants, likely by transient increase in structural stability upon antagonist binding. Based on this premise, we examined whether antagonists could enhance OR membrane expression in heterologous cells. Using phenyl salicylate (PES) on cells expressing the mouse OR Or11g7, we observed increased cell surface expression exceeding the effects of co-expression with the OR chaperone RTP1S. After removing the antagonist, Or11g7 retained normal agonist responsiveness. Similar enhancements in cell surface expression were observed for a human OR OR2T11 treated with its antagonists. These findings suggest that small-molecule antagonists act as pharmacological chaperones to stabilize OR conformation, enhancing surface expression in a manner similar to molecular chaperones. Our study reveals a novel role for odorant antagonists in OR biogenesis and may inform future research on olfactory training mechanisms.