title: Agonist-induced long-term desensitization of the human prostacyclin receptor.
abstract: Phosphorylation of the human prostacyclin (PGI(2)) receptor (hIP-R) by diacylglycerol-regulated protein kinase C (PKC) has been reported to be responsible for its rapid desensitization in HEK293 cells. In this study we demonstrate, that human fibroblasts reveal a much slower hIP-R desensitization kinetics, which was neither affected by stimulation nor inhibition of PKC by either phorbol 12-myristate-13-acetate or GF-109203X suggesting a different cellular mechanism. Although agonist-promoted sequestration of a C-terminally green fluorescent protein-tagged hIP-R was demonstrated, it did not account for the long-term desensitization. Concanavalin A did not abolish, but accelerated receptor desensitization kinetics. Resensitization of hIP-R involved receptor recycling and/or de novo synthesis of receptor protein, depending on the duration of prior desensitization. This is the first study investigating the mechanisms of hIP-R desensitization in intact human cells naturally expressing hIP-R. Our data suggest, that a hitherto unknown mechanism of hIP-R long-term desensitization, which is independent of receptor phosphorylation by conventional and novel type PKC isoforms or endocytosis, is a key event in regulating the cellular responsiveness to PGI(2).
authors.affiliation: Institut für Pharmakologie und klinische Pharmakologie, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: S M
authors.initials: SM
authors.lastname: Nilius
authors.forename: A
authors.initials: A
authors.lastname: Hasse
authors.forename: P
authors.initials: P
authors.lastname: Kuger
authors.forename: K
authors.initials: K
authors.lastname: Schrör
authors.forename: J
authors.initials: J
authors.lastname: Meyer-Kirchrath
mesh_terms: D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D054774:G-Protein-Coupled Receptor Kinase 5; D006801:Humans; D016285:Iloprost / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology; D007700:Kinetics; D008301:Maleimides / Q000494:pharmacology; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D051552:beta-Adrenergic Receptor Kinases
chemical_list: D007211:Indoles; D008301:Maleimides; D044006:Receptors, Epoprostenol; D011982:Receptors, Prostaglandin; D011993:Recombinant Fusion Proteins; D000242:Cyclic AMP; D017346:Protein Serine-Threonine Kinases; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D051552:beta-Adrenergic Receptor Kinases; D054774:G-Protein-Coupled Receptor Kinase 5; C517898:GRK5 protein, human; D016285:Iloprost; C070515:bisindolylmaleimide I; D013755:Tetradecanoylphorbol Acetate