scholarly journals Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

10.3791/55514 ◽  
2017 ◽  
Author(s):  
Jingwen Yang ◽  
Yunjun Yan ◽  
Xiaowei Xiang ◽  
Yuchao Xu ◽  
Naiming Zhou ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
G Protein ◽  
Receptor Internalization ◽  
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 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.

scholarly journals Dynamin and β-Arrestin Reveal Distinct Mechanisms for G Protein-coupled Receptor Internalization

1996 ◽  
Vol 271 (31) ◽  
pp. 18302-18305 ◽  
Author(s):  
Jie Zhang ◽  
Stephen S. G. Ferguson ◽  
Larry S. Barak ◽  
Luc Ménard ◽  
Marc G. Caron
Keyword(s):  
G Protein ◽  
Receptor Internalization ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

A novel enzyme complementation‐based assay for monitoring G‐protein‐coupled receptor internalization

2007 ◽  
Vol 21 (14) ◽  
pp. 3827-3834 ◽  
Author(s):  
Mark M. Hammer ◽  
Tom S. Wehrman ◽  
Helen M. Blau
Keyword(s):  
G Protein ◽  
Receptor Internalization ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled

Role of beta-Arrestin in Mediating Agonist-Promoted G Protein-Coupled Receptor Internalization

Science ◽  
1996 ◽  
Vol 271 (5247) ◽  
pp. 363-366 ◽  
Author(s):  
S. S. G. Ferguson ◽  
W. E. Downey ◽  
A.-M. Colapietro ◽  
L. S. Barak ◽  
L. M nard ◽  
...  
Keyword(s):  
G Protein ◽  
Receptor Internalization ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled
Sign in / Sign up

Export Citation Format

Share Document