Palmitoylation of the rat μ opioid receptor

FEBS Letters

Shahabi, Vafa¹  Liu-Chen, Lee-Yuan¹  Chen, Chongguang¹  Xu, Wei¹ 

[1] Department of Pharmacology, Temple University School of Medicine, 3420 N. Broad St., Philadelphia, PA 19140, USA

关键词: Palmitoylation;    Mu opioid receptor;    CHO cells;    Chinese hamster ovary cells;    CHO-μR cells;    Chinese hamster ovary cells stably expressing the rat μ opioid receptor;    DAMGO;    Try-d-Ala-Gly-N-Me-Phe-Gly-ol;    TLC;    thin layer chromatography;    GPCR;    G-protein coupled receptor;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;   

DOI  :  10.1016/S0014-5793(98)01547-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We examined whether the μ opioid receptor was palmitoylated and attempted to determine sites of palmitoylation. Following metabolic labeling with [³H]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 [³H]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 [³H]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.

【 授权许可】

Unknown