Sixty percent of all cloned human G protein-coupled receptors (GPCRs) have no identifiable agonist, so are termed �orphan receptors.� The deduced primary structure of the human orphan GPCR, GP133, is similar to four previously identified D. discoideum GPCRs whose agonist, cAMP, initiates both chemotaxis and cellular development. This suggests that the agonist of GP133 may also be cAMP. A consensus C-terminal, cytoplasmic PDZ domain may promote GP133 expression by interacting with membrane scaffold proteins. Green fluorescent protein (GFP) cDNA has been inserted C-terminal to the GP133 cDNA to form a continuous open reading frame and thus provide a visual expression marker. A second construct duplicates the first with the addition of a second GP133 PDZ domain C-terminal to the GFP cDNA. Both fusion proteins will be expressed in mammalian cells to determine cAMP binding, the intracellular signaling pathway of GP133 and the necessity of a C-terminal PDZ domain.

Cell surface expression and characterization of a human orphan G protein-coupled receptor

243rd National Meeting (2012)

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