Determining which domains and amino acid residues of the μ opioid receptor are phosphorylated is critical for understanding the mechanism of μ opioid receptor phosphorylation. The role of the C-terminus of the receptor was investigated by examining the C-terminally truncated or point-mutated μ opioid receptors in receptor phosphorylation and desensitization. Both wild- type and mutated receptors were stably expressed in Chinese hamster ovary (CHO) cells. The receptor expression was confirmed by receptor radioligand binding and immunoblottting. After exposure to 5 μM of DAMGO, phosphorylation of the C-terminally truncated receptor and the mutant receptor T394A was reduced to 40 and 10% of that of the wild-type receptor, respectively. Mutation effects on agonist-induced desensitization were studied using adenylyl cyclase inhibition assays. The C-terminally truncated receptor and mutant receptor T394A both showed complete loss of DAMGO-induced desensitization, while the mutant T/S7A receptor only lost part of its ability to desensitize. Taken together, these results suggest that the C- terminus of the μ opioid receptor participates in receptor phosphorylation and desensitization with threonine 394, a crucial residue for both features. DAMGO-induced μ opioid receptor phosphorylation and desensitization are associated and appear to involve both the μ opioid receptor C-terminus and other domains of the receptor.
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