Elimination of palmitoylation sites in the human dopamine D1 receptor does not affect receptor-G protein interaction.
We have eliminated putative palmitoylation sites in the carboxyl tail of the human dopamine D1 receptor by replacing the two cysteine residues with alanines either separately or together. The wild type and the three mutated dopamine D1 receptors were stably expressed in baby hamster kidney cells and characterized to detect any resulting alterations in receptor-G protein interactions. The three mutant dopamine D1 receptors retained the same proportion of high affinity state for agonists as wild type receptors and also no difference was observed in the stimulation of adenylyl cyclase activity. Our results are in contrast to those observed with the beta 2-adrenoceptor and consistent with similar studies of luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptors, alpha 2-adrenoceptors, muscarinic M2 receptors and thyrotropin releasing hormone (TRH) receptors. Thus, we suggest that palmitoylation appears to play a unique role in the beta 2-adrenoceptors, and appears not to be essential in G protein coupling for the dopamine D1 receptors.
Jin
H
H
Department of Pharmacology, University of Toronto, Ontario, Canada.
Zastawny
R
R
George
S R
SR
O'Dowd
B F
BF
D015647:2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine / Q000378:metabolism / Q000494:pharmacology; D000215:Acylation; D000262:Adenylyl Cyclases / Q000378:metabolism; D000409:Alanine / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D006224:Cricetinae; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism; D018491:Dopamine Agonists / Q000378:metabolism / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019308:Palmitic Acid / Q000378:metabolism; D011869:Radioligand Assay; D018343:Receptors, Adrenergic, beta-2 / Q000187:drug effects* / Q000378:metabolism; D017447:Receptors, Dopamine D1 / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment; D012984:Software; D014162:Transfection
D018491:Dopamine Agonists; D018343:Receptors, Adrenergic, beta-2; D017447:Receptors, Dopamine D1; D019308:Palmitic Acid; D015647:2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D003545:Cysteine; D000409:Alanine; D004298:Dopamine