title: Rapid agonist-induced phosphorylation of the human CRF receptor, type 1: a potential mechanism for homologous desensitization.
abstract: Agonist-induced phosphorylation of the human corticotropin-releasing factor type 1 receptor (hCRF(1)-R) was investigated using an influenza hemagglutinin (HA) epitope-tagged receptor transiently expressed in COS-7 cells. The HA-hCRF(1)-R migrated as a broad band (M(r) 60,000-70,000) in SDS-PAGE and showed increased mobility (M(r) approximately 48,000) after enzymatic deglycosylation with peptide-N-glycosidase F, consistent with the predicted size (47 kDa) of the nonglycosylated HA-hCRF(1)-R protein. A marked increase in HA-hCRF(1)-R phosphorylation was observed in HA-hCRF(1)-R-expressing COS-7 cells exposed to 1 microM ovine CRF for 5 min, whereas activation of protein kinase A (PKA) by 50 microM forskolin, or of Ca(2+)/calmodulin (CaM)-dependent kinases by 10 microM ionomycin, had little effect. These findings are consistent with preliminary data suggesting that CRF(1)-R phosphorylation mediated by G protein receptor kinase 3 (GRK3), but not by PKA or CaM-dependent kinases, has an important role in the homologous desensitization of brain CRF(1)-Rs.
authors.affiliation: VA Medical Center and Department of Psychiatry, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0603, USA. rhauger@ucsd.edu
authors.forename: R L
authors.initials: RL
authors.lastname: Hauger
authors.forename: R D
authors.initials: RD
authors.lastname: Smith
authors.forename: S
authors.initials: S
authors.lastname: Braun
authors.forename: F M
authors.initials: FM
authors.lastname: Dautzenberg
authors.forename: K J
authors.initials: KJ
authors.lastname: Catt
mesh_terms: D000818:Animals; D019556:COS Cells; D003346:Corticotropin-Releasing Hormone / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D018019:Receptors, Corticotropin-Releasing Hormone / Q000819:agonists / Q000378:metabolism*; D000099052:CRF Receptor, Type 1
chemical_list: D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1; D003346:Corticotropin-Releasing Hormone