Expression cloning of cDNA encoding a seven-helix receptor from human placenta with affinity for opioid ligands

Abstract

Here we report the expression cloning of cDNA encoding a putative opioid receptor from a human placenta cDNA library. Placental opioid receptors are of the κ type. As the dynorphin opioid peptides are κ-selective, a dynorphin ligand was used in an affinity-enrichment (panning) procedure to select transiently transfected COS-7 cells expressing κ receptor binding sites. The cloned cDNA encodes a 440-residue protein of the seven-helix guanine nucleotide-binding protein (G-protein)-coupled receptor family. Ligand binding reveals a stereospecific site with typical opioid properties, which binds peptide and nonpeptide opioids with moderate affinity (Kd ≈ 100 nM) and which lacks the expected K selectivity. The deduced transmembrane domain is 93% identical to the homologous region of the human neuromedin K (neurokinin B) receptor, but the N-terminal and C-terminal sequences have many dissimilarities. The expressed receptor binds opioid ligands but not tachykinins; and under the same conditions, a cloned rat neuromedin K receptor binds tachykinins but not opioids.