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Palmitoylation of the rat μ opioid receptor
Author(s) -
Chen Chongguang,
Shahabi Vafa,
Xu Wei,
Liu-Chen Lee-Yuan
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)01547-6
Subject(s) - palmitoylation , palmitic acid , receptor , chemistry , opioid receptor , transfection , dithiothreitol , μ opioid receptor , chinese hamster ovary cell , hek 293 cells , biochemistry , microbiology and biotechnology , cysteine , opioid , biology , fatty acid , enzyme , gene
We examined whether the μ opioid receptor was palmitoylated and attempted to determine sites of palmitoylation. Following metabolic labeling with [ 3 H]palmitic acid and immunoaffinity purification of the μ opioid receptor, SDS‐PAGE and fluorography revealed a broad labeled band with M r of ∼80 kDa in CHO cells stably expressing the rat μ receptor, but not in CHO cells transfected with the vector alone, indicating that the μ receptor is palmitoylated. Activation of the receptor with morphine did not affect the extent of palmitoylation. Hydroxylamine or dithiothreitol treatment removed most of the radioactivity, demonstrating that [ 3 H]palmitic acid is incorporated into Cys residue(s) via thioester bond(s). Surprisingly, mutations of the only two Cys residues in the C‐terminal domain did not reduce [ 3 H]palmitic acid incorporation significantly. Thus, unlike many G‐protein coupled receptors, the palmitoylation site(s) of the rat μ opioid receptor do(es) not reside in the C‐terminal domain.