scholarly journals Phosphorylation Analysis of G Protein-Coupled Receptor by Mass Spectrometry: Identification of a Phosphorylation Site in V2 Vasopressin Receptor

2008 ◽  
Vol 80 (15) ◽  
pp. 6034-6037 ◽  
Author(s):  
Shilan Wu ◽  
Mariel Birnbaumer ◽  
Ziqiang Guan
Keyword(s):  
Mass Spectrometry ◽  
G Protein ◽  
Phosphorylation Site ◽  
Vasopressin Receptor ◽  
G Protein Coupled Receptor ◽  
Mass Spectrometry Identification ◽  
V2 Vasopressin Receptor ◽  
G Protein Coupled ◽  
Phosphorylation Analysis

Analysis of an Intact G-Protein Coupled Receptor by MALDI-TOF Mass Spectrometry:  Molecular Heterogeneity of the Tachykinin NK-1 Receptor

2007 ◽  
Vol 79 (6) ◽  
pp. 2189-2198 ◽  
Author(s):  
Isabel D. Alves ◽  
Emmanuelle Sachon ◽  
Gerard Bolbach ◽  
Lynda Millstine ◽  
Solange Lavielle ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
G Protein ◽  
Molecular Heterogeneity ◽  
G Protein Coupled Receptor ◽  
Maldi Tof Mass Spectrometry ◽  
Maldi Tof ◽  
G Protein Coupled

scholarly journals Accelerating the Throughput of Affinity Mass Spectrometry-Based Ligand Screening toward a G Protein-Coupled Receptor

2019 ◽  
Vol 91 (13) ◽  
pp. 8162-8169 ◽  
Author(s):  
Yan Lu ◽  
Shanshan Qin ◽  
Bingjie Zhang ◽  
Antao Dai ◽  
Xiaoqing Cai ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
G Protein ◽  
G Protein Coupled Receptor ◽  
Ligand Screening ◽  
G Protein Coupled

scholarly journals G Protein-coupled Receptor Kinase 2–mediated Phosphorylation of Ezrin Is Required for G Protein-coupled Receptor–dependent Reorganization of the Actin Cytoskeleton

2005 ◽  
Vol 16 (7) ◽  
pp. 3088-3099 ◽  
Author(s):  
Sarah H. Cant ◽  
Julie A. Pitcher
Keyword(s):  
G Protein ◽  
Phosphorylation Site ◽  
Rho Kinase ◽  
Receptor Kinase ◽  
G Protein Coupled Receptor ◽  
Glutathione S Transferase ◽  
Membrane Ruffling ◽  
Gpcr Activation ◽  
G Protein Coupled

G protein-coupled receptor kinase 2 (GRK2) phosphorylates and desensitizes activated G protein-coupled receptors (GPCRs). Here, we identify ezrin as a novel non-GPCR substrate of GRK2. GRK2 phosphorylates glutathione S-transferase (GST)-ezrin, but not an ezrin fusion protein lacking threonine 567 (T567), in vitro. These results suggest that T567, the regulatory phosphorylation site responsible for maintaining ezrin in its active conformation, represents the principle site of GRK2-mediated phosphorylation. Two lines of evidence indicate that GRK2-mediated ezrin-radixinmoesin (ERM) phosphorylation serves to link GPCR activation to cytoskeletal reorganization. First, in Hep2 cells muscarinic M1 receptor (M1MR) activation causes membrane ruffling. This ruffling response is ERM dependent and is accompanied by ERM phosphorylation. Inhibition of GRK2, but not rho kinase or protein kinase C, prevents ERM phosphorylation and membrane ruffling. Second, agonist-induced internalization of the β2-adrenergic receptor (β2AR) and M1MR is accompanied by ERM phosphorylation and localization of phosphorylated ERM to receptor-containing endocytic vesicles. The colocalization of internalized β2AR and phosphorylated ERM is not dependent on Na+/H+ exchanger regulatory factor binding to the β2AR. Inhibition of ezrin function impedes β2AR internalization, further linking GPCR activation, GRK activity, and ezrin function. Overall, our results suggest that GRK2 serves not only to attenuate but also to transduce GPCR-mediated signals.

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 High-throughput identification of G protein-coupled receptor modulators through affinity mass spectrometry screening

2018 ◽  
Vol 9 (12) ◽  
pp. 3192-3199 ◽  
Author(s):  
Shanshan Qin ◽  
Mengmeng Meng ◽  
Dehua Yang ◽  
Wenwen Bai ◽  
Yan Lu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
G Protein ◽  
High Throughput ◽  
G Protein Coupled Receptor ◽  
Receptor Modulators ◽  
G Protein Coupled

High-throughput identification of GPCR modulators through affinity MS screening.

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