Phosphorylation and palmitoylation of the human D2L dopamine receptor in Sf9 cells.
We have expressed and biochemically characterized the human D2long (D2L) dopamine receptor isoform using the baculovirus/Sf9 cell system. The expressed receptor bound ligands with a pharmacological profile similar to that reported for neuronal and cloned D2L receptors expressed in mammalian cell lines. Dopamine binding to D2L receptor was sensitive to guanine nucleotides, indicating receptor coupling to endogenous G proteins. A D2L receptor-specific antibody identified two major protein species at approximately 44 kDa and at approximately 93 kDa in immunoblots, suggesting the presence of D2L receptor monomers and dimers. Both species were purified by immunoprecipitation from digitonin-solubilized preparation of cells expressing D2L receptor prelabeled with 32P(i) or [3H]-palmitate. These results constitute the first direct evidence for D2L receptor phosphorylation and palmitoylation.
Ng
G Y
GY
Department of Pharmacology, University of Toronto, Ontario, Canada.
O'Dowd
B F
BF
Caron
M
M
Dennis
M
M
Brann
M R
MR
George
S R
SR
D000818:Animals; D002460:Cell Line; D004298:Dopamine / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000032:analysis / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007536:Isomerism; D009036:Moths / Q000166:cytology*; D010168:Palmitates / Q000032:analysis / Q000378:metabolism*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011233:Precipitin Tests; D017448:Receptors, Dopamine D2 / Q000032:analysis / Q000378:metabolism* / Q000502:physiology; D014316:Tritium
D010168:Palmitates; D010761:Phosphorus Radioisotopes; D017448:Receptors, Dopamine D2; D014316:Tritium; D019204:GTP-Binding Proteins; D004298:Dopamine