Eliminating phosphorylation sites of the parathyroid hormone receptor type 1 differentially affects stimulation of phospholipase C and receptor internalization.
The parathyroid hormone (PTH)/PTH-related peptide (PTHrP) receptor (PTH1R) belongs to family B of seven-transmembrane-spanning receptors and is activated by PTH and PTHrP. Upon PTH stimulation, the rat PTH1R becomes phosphorylated at seven serine residues. Elimination of all PTH1R phosphorylation sites results in prolonged cAMP accumulation and impaired internalization in stably transfected LLC-PK1 cells. The present study explores the role of individual PTH1R phosphorylation sites in PTH1R signaling through phospholipase C, agonist-dependent receptor internalization, and regulation by G protein-coupled receptor kinases. By means of transiently transfected COS-7 cells, we demonstrate that the phosphorylation-deficient (pd) PTH1R confers dramatically enhanced coupling to G(q/11) proteins upon PTH stimulation predominantly caused by elimination of Ser(491/492/493), Ser(501), or Ser(504). Reportedly, impaired internalization of the pd PTH1R, however, is not dependent on a specific phosphorylation site. In addition, we show that G protein-coupled receptor kinase 2 interferes with pd PTH1R signaling to G(q/11) proteins at least partially by direct binding to G(q/11) proteins.
Miedlich
Susanne U
SU
Massachusetts General Hospital/Harvard Medical School, Boston, MA 02114, USA. smiedlich@partners.org
Abou-Samra
Abdul B
AB
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003627:Data Interpretation, Statistical; D054768:G-Protein-Coupled Receptor Kinases / Q000378:metabolism; D007295:Inositol Phosphates / Q000378:metabolism; D018374:LLC-PK1 Cells; D009154:Mutation / Q000235:genetics / Q000502:physiology; D010281:Parathyroid Hormone / Q000378:metabolism; D010766:Phosphorylation; D010957:Plasmids; D051381:Rats; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000187:drug effects; D012694:Serine / Q000737:chemistry; D013552:Swine; D014162:Transfection; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism*
D007295:Inositol Phosphates; D010281:Parathyroid Hormone; D044168:Receptor, Parathyroid Hormone, Type 1; D011956:Receptors, Cell Surface; D012694:Serine; D054768:G-Protein-Coupled Receptor Kinases; D010738:Type C Phospholipases