Sixty percent of all human G protein-coupled receptors (GPCRs), including GPR133, do not have an identified agonist, so are termed �orphan receptors.� The primary structure of GPR133 is similar to four previously identified D. discoideum GPCRs whose agonist, extracellular cAMP, initiates chemotaxis and cellular development suggesting that the agonist of GPR133 may also be cAMP. To ensure high expression levels, a mammalian expression vector was constructed that included the sequence encoding the N-terminal 20 amino acids of bovine rhodopsin inserted after the signal peptide sequence of GPR133. After transfection of COS-7 cells, the ability of extracellular cAMP to increase intracellular cAMP in a GPR133-dependent fashion was assessed using a fluorescence resonance energy transfer assay.

Expression and identification of GPR133, a human orphan G-protein coupled receptor

245th National Meeting (2013)

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