Measuring Cell Surface Expression of Odorant Receptors via Flow Cytometry.

Odorant receptor proteins (ORs) have highly variable cell surface expression levels. The majority of both human and murine ORs depend on chaperone proteins to traffic from the endoplasmic reticulum to the cell surface, while a limited subset of ORs express at high levels independently. Quantifying these heterogeneous expression levels is of high import for understanding the trafficking and stability of these integral-transmembrane proteins and for normalizing in vitro activation assays. Recognizable epitopes like the rhodopsin-tag can be inserted upstream of the N-termini in ORs to enable cell surface immunostaining and detection via flow cytometry. This method enables robust measurement and comparison of cell surface expression levels of different ORs. Our approach also facilitates the study of different chaperone proteins' effects on OR trafficking and expression.

Duke Scholars

Author Hiroaki Matsunami Molecular Genetics and Microbiology