scholarly journals Eliminating phosphorylation sites of the parathyroid hormone receptor type 1 differentially affects stimulation of phospholipase C and receptor internalization

2008 ◽  
Vol 295 (3) ◽  
pp. E665-E671 ◽  
Author(s):  
Susanne U. Miedlich ◽  
Abdul B. Abou-Samra
Keyword(s):  
Parathyroid Hormone ◽  
Phospholipase C ◽  
G Protein ◽  
Phosphorylation Site ◽  
Receptor Internalization ◽  
Phosphorylation Sites ◽  
G Protein Coupled Receptor ◽  
Parathyroid Hormone Receptor ◽  
Direct Binding ◽  
G Protein Coupled

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 Gq/11 proteins upon PTH stimulation predominantly caused by elimination of Ser491/492/493, Ser501, or Ser504. 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 Gq/11 proteins at least partially by direct binding to Gq/11 proteins.

scholarly journals Apoptosis Mediated by Activation of the G Protein-Coupled Receptor for Parathyroid Hormone (PTH)/ PTH-Related Protein (PTHrP)

2000 ◽  
Vol 14 (2) ◽  
pp. 241-254 ◽  
Author(s):  
Paul R. Turner ◽  
Suzanne Mefford ◽  
Sylvia Christakos ◽  
Robert A. Nissenson
Keyword(s):  
Parathyroid Hormone ◽  
G Protein ◽  
Related Protein ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

scholarly journals Distinct phosphorylation sites in a prototypical GPCR differently orchestrate β-arrestin interaction, trafficking, and signaling

2020 ◽  
Vol 6 (37) ◽  
pp. eabb8368 ◽  
Author(s):  
Hemlata Dwivedi-Agnihotri ◽  
Madhu Chaturvedi ◽  
Mithu Baidya ◽  
Tomasz Maciej Stepniewski ◽  
Shubhi Pandey ◽  
...  
Keyword(s):  
G Protein ◽  
Phosphorylation Site ◽  
Dynamics Simulation ◽  
Phosphorylation Sites ◽  
Functional Responses ◽  
Biased Signaling ◽  
The Stability ◽  
G Protein Coupled ◽  
Structural Aspects

Agonist-induced phosphorylation of G protein–coupled receptors (GPCRs) is a key determinant for their interaction with β-arrestins (βarrs) and subsequent functional responses. Therefore, it is important to decipher the contribution and interplay of different receptor phosphorylation sites in governing βarr interaction and functional outcomes. Here, we find that several phosphorylation sites in the human vasopressin receptor (V2R), positioned either individually or in clusters, differentially contribute to βarr recruitment, trafficking, and ERK1/2 activation. Even a single phosphorylation site in V2R, suitably positioned to cross-talk with a key residue in βarrs, has a decisive contribution in βarr recruitment, and its mutation results in strong G-protein bias. Molecular dynamics simulation provides mechanistic insights into the pivotal role of this key phosphorylation site in governing the stability of βarr interaction and regulating the interdomain rotation in βarrs. Our findings uncover important structural aspects to better understand the framework of GPCR-βarr interaction and biased signaling.

scholarly journals Cell Density Sensing Mediated by a G Protein-coupled Receptor Activating Phospholipase C

1998 ◽  
Vol 273 (14) ◽  
pp. 8161-8168 ◽  
Author(s):  
Derrick T. Brazill ◽  
David F. Lindsey ◽  
John D. Bishop ◽  
Richard H. Gomer
Keyword(s):  
Phospholipase C ◽  
G Protein ◽  
Cell Density ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

scholarly journals A pH-Sensitive Fluor, CypHerTM 5, Used to Monitor Agonist-Induced G Protein-Coupled Receptor Internalization in Live Cells

BioTechniques ◽  
2002 ◽  
Vol 33 (5) ◽  
pp. 1152-1157 ◽  
Author(s):  
E.J. Adie ◽  
S. Kalinka ◽  
L. Smith ◽  
M.J. Francis ◽  
A. Marenghi ◽  
...  
Keyword(s):  
G Protein ◽  
Ph Sensitive ◽  
Receptor Internalization ◽  
Live Cells ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

scholarly journals WDR36 acts as a scaffold protein tethering a G-protein-coupled receptor, G q and phospholipase C  in a signalling complex

2011 ◽  
Vol 124 (19) ◽  
pp. 3292-3304 ◽  
Author(s):  
A. Cartier ◽  
A. Parent ◽  
P. Labrecque ◽  
G. Laroche ◽  
J.-L. Parent
Keyword(s):  
Phospholipase C ◽  
G Protein ◽  
Scaffold Protein ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

scholarly journals Role of G Protein-Coupled Receptor Kinases on the Agonist-Induced Phosphorylation and Internalization of the Follitropin Receptor

1999 ◽  
Vol 13 (6) ◽  
pp. 866-878 ◽  
Author(s):  
Maria de Fatima M. Lazari ◽  
Xuebo Liu ◽  
Kazuto Nakamura ◽  
Jeffrey L. Benovic ◽  
Mario Ascoli
Keyword(s):  
Cell Line ◽  
G Protein ◽  
Dominant Negative ◽  
Receptor Internalization ◽  
Dominant Negative Mutant ◽  
Deficient Mutant ◽  
G Protein Coupled Receptor ◽  
Western Blots ◽  
293 Cells ◽  
G Protein Coupled

Abstract The experiments presented herein were designed to identify members of the G protein-coupled receptor kinase (GRK) family that participate in the agonist-induced phosphorylation and internalization of the rat FSH receptor (rFSHR). Western blots of human kidney 293 cells (the cell line used in transfection experiments) and MSC-1 cells (a cell line derived from Sertoli cells that displays many of the differentiated functions of their normal counterparts) reveal the presence of GRK2 and GRK6 in both cell lines as well as GRK4 in MSC-1 cells. Cotransfection of 293 cells with the rFSHR and GRK2, GRK4α, or GRK6 resulted in an increase in the agonist-induced phosphorylation of the rFSHR. Cotransfections of the rFSHR with GRKs or arrestin-3 enhanced the agonist-induced internalization of the rFHSR, and combinations of GRKs and arrestin-3 were more effective than the individual components. To characterize the involvement of endogenous GRKs on phosphorylation and internalization, we inhibited endogenous GRK2 by overexpression of a kinase-deficient mutant of GRK2 or Gαt, a scavenger of Gβγ. We also inhibited endogenous GRK6 by overexpression of a kinase-deficient mutant of GKR6. All three constructs were effective inhibitors of phosphorylation, but only the kinase-deficient mutant of GRK2 and Gαt inhibited internalization. The inhibition of internalization induced by these two constructs was less pronounced than that induced by a dominant-negative mutant of the nonvisual arrrestins, however. The finding that inhibitors of GRK2 and GRK6 impair phosphorylation, but only the inhibitors of GRK2 impair internalization, suggests that different GRKs have differential effects on receptor internalization.

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