title: Palmitoylation of human thyrotropin receptor: slower intracellular trafficking of the palmitoylation-defective mutant.
abstract: We here show that the epitope-tagged human TSH receptor (TSHRmyc) is covalently modified with palmitic acid by thioesterification. Side-directed mutagenesis identified Cys699 in the C-terminal cytoplasmic tail of the receptor as the putative palmitoylation site. Mutation of Cys699 to Ala results in the nonpalmitoylated receptor (TSHRmycC699A) in which high affinity TSH binding, Gs coupling, homologous desensitization and TSH-induced internalization are unaffected. In contrast, abolition of palmitoylation appears to decrease the rate of the intracellular trafficking of the receptor. However, since most of TSHRmycC699A seems to be fully processed finally and the receptor number of TSHRmycC699A on the cell surface is comparable to that of TSHRmyc, our results suggest that abolition of palmitoylation delays the cell surface expression of TSHR, but does not trap the receptor intracellularly, although another possibility for proteolytic degradation of either the 95 kDa or the 100 kDa mutant receptor can not be excluded. Thus, post-translational modification of TSHR by palmitoylation may provide a novel mechanism of enhancing the rate of intracellular trafficking of the receptor.
authors.affiliation: Department of Pharmacology 1, Atomic Bomb Disease Institute, Nagasaki University School of Medicine, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Tanaka
authors.forename: Y
authors.initials: Y
authors.lastname: Nagayama
authors.forename: E
authors.initials: E
authors.lastname: Nishihara
authors.forename: H
authors.initials: H
authors.lastname: Namba
authors.forename: S
authors.initials: S
authors.lastname: Yamashita
authors.forename: M
authors.initials: M
authors.lastname: Niwa
mesh_terms: D000595:Amino Acid Sequence; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000502:physiology*; D019308:Palmitic Acid / Q000378:metabolism*; D011989:Receptors, Thyrotropin / Q000235:genetics / Q000378:metabolism*; D016324:Sequence Tagged Sites
chemical_list: D011989:Receptors, Thyrotropin; D019308:Palmitic Acid