Received for publication June 6, 2002.
Revised July 19, 2002.
Accepted for publication November 18, 2002.

Serine 329 of the mu opioid receptor interacts differently with agonists

Abstract

To investigate the effect of the hydrophilic Ser amino acid in position 329 of the human µ-opioid receptor (hMORwt) on the potency of various agonists, we mutated this residue to Ala (hMORS329A). Taking advantage of the functional coupling of the opioid receptor with the heteromultimeric G-protein-coupled inwardly rectifying potassium channel (GIRK1/GIRK2), either the wild type hMOR or the mutated receptor (hMORS329A) was functionally coexpressed with GIRK1- and GIRK2- channels together with a regulator of G-protein signalling (RGS4) in Xenopus laevis oocytes. The two-microelectrode voltage clamp technique was used to measure the opioid receptor activated GIRK1/GIRK2 channel responses. The potency of the peptide agonist DAMGO decreased as measured via hMORS329A, while the potency of non peptide agonists like morphine, fentanyl and -hydroxyfentanyl (R004333) increased via the mutated receptor. Our results are indicative for the existence of hydrophilic interactions between Ser329 and DAMGO, thereby decreasing the potency of DAMGO via the mutated receptor, while hydrophobic interactions between the mutated receptor and the N-phenylethyl of morphine and fentanyl can explain the increased potency. We conclude that the hydroxyl group of Ser329 is not involved in the formation of a hydrogen bond with the -hydroxy group of fentanyl and that mutation of this residue to alanine caused dual effects depending on the nature of the ligand.

Key words: cloned opioid receptors, fentanyl, pain, rgs, voltage clamp, xenopus oocytes