title: Palmitoylation occurs at cysteine 347 and cysteine 351 of the dopamine D(1) receptor.
abstract: To determine the palmitoylation sites in the human dopamine D(1) receptor, we expressed wild type and mutant receptors in which candidate cysteines in the carboxyl tail were substituted by alanines both individually (A347, A351) and together (AA). Our results showed that palmitoylation levels of A347 and A351 were reduced substantially and that AA had no detectable signal of palmitoylation. These data indicate that cysteines 347 and 351 were both palmitoylated and that they were the only sites of palmitoylation. We introduced a cAMP-dependent protein kinase site encompassing the position 351. We predicted that a functional cAMP-dependent protein kinase site would impair receptor-G protein coupling if it is not occluded by palmitoylation. Our results demonstrated that indeed, the introduction of the cAMP-dependent protein kinase site caused reduced potency of dopamine stimulation of adenylyl cyclase, and thus confirmed that when unoccluded, the cAMP-dependent protein kinase site introduced to position 351 of dopamine D(1) receptor could confer constitutive desensitization.
authors.affiliation: Department of Pharmacology, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Jin
authors.forename: Z
authors.initials: Z
authors.lastname: Xie
authors.forename: S R
authors.initials: SR
authors.lastname: George
authors.forename: B F
authors.initials: BF
authors.lastname: O'Dowd
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D000242:Cyclic AMP / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D007313:Insecta; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D017447:Receptors, Dopamine D1 / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D018343:Receptors, Adrenergic, beta-2; D017447:Receptors, Dopamine D1; D000242:Cyclic AMP; D011494:Protein Kinases; D019204:GTP-Binding Proteins; D003545:Cysteine