scholarly journals Plasma Membrane and Nuclear Localization of G Protein–coupled Receptor Kinase 6A

2007 ◽  
Vol 18 (8) ◽  
pp. 2960-2969 ◽  
Author(s):  
Xiaoshan Jiang ◽  
Jeffrey L. Benovic ◽  
Philip B. Wedegaertner
Keyword(s):  
Amino Acids ◽  
Plasma Membrane ◽  
G Protein ◽  
Nuclear Localization ◽  
Nuclear Export ◽  
Amphipathic Helix ◽  
G Protein Coupled Receptor ◽  
C Terminus ◽  
Acidic Residues ◽  
G Protein Coupled

G protein–coupled receptor (GPCR) kinases (GRKs) specifically phosphorylate agonist-occupied GPCRs at the inner surface of the plasma membrane (PM), leading to receptor desensitization. Here we show that the C-terminal 30 amino acids of GRK6A contain multiple elements that either promote or inhibit PM localization. Disruption of palmitoylation by individual mutation of cysteine 561, 562, or 565 or treatment of cells with 2-bromopalmitate shifts GRK6A from the PM to both the cytoplasm and nucleus. Likewise, disruption of the hydrophobic nature of a predicted amphipathic helix by mutation of two leucines to alanines at positions 551 and 552 causes a loss of PM localization. Moreover, acidic amino acids in the C-terminus appear to negatively regulate PM localization; mutational replacement of several acidic residues with neutral or basic residues rescues PM localization of a palmitoylation-defective GRK6A. Last, we characterize the novel nuclear localization, showing that nuclear export of nonpalmitoylated GRK6A is sensitive to leptomycin B and that GRK6A contains a potential nuclear localization signal. Our results suggest that the C-terminus of GRK6A contains a novel electrostatic palmitoyl switch in which acidic residues weaken the membrane-binding strength of the amphipathic helix, thus allowing changes in palmitoylation to regulate PM versus cytoplasmic/nuclear localization.

scholarly journals G Protein-Coupled Receptor Kinase 5 Contains a DNA-Binding Nuclear Localization Sequence

2004 ◽  
Vol 24 (23) ◽  
pp. 10169-10179 ◽  
Author(s):  
Laura R. Johnson ◽  
Mark G. H. Scott ◽  
Julie A. Pitcher
Keyword(s):  
Dna Binding ◽  
G Protein ◽  
Nuclear Localization ◽  
Nuclear Export ◽  
Calcium Ionophore ◽  
Nuclear Localization Sequence ◽  
G Protein Coupled Receptor ◽  
Gpcr Activation ◽  
Localization Sequence ◽  
G Protein Coupled

ABSTRACT G protein-coupled receptor kinases (GRKs) mediate desensitization of agonist-occupied G protein-coupled receptors (GPCRs). Here we report that GRK5 contains a DNA-binding nuclear localization sequence (NLS) and that its nuclear localization is regulated by GPCR activation, results that suggest potential nuclear functions for GRK5. As assessed by fluorescence confocal microscopy, transfected and endogenous GRK5 is present in the nuclei of HEp2 cells. Mutation of basic residues in the catalytic domain of GRK5 (between amino acids 388 and 395) results in the nuclear exclusion of the mutant enzyme (GRK5Δ NLS), demonstrating that GRK5 contains a functional NLS. The nuclear localization of GRK5 is subject to dynamic regulation. Calcium ionophore treatment or activation of Gq-coupled muscarinic-M3 receptors promotes the nuclear export of the kinase in a Ca2+/calmodulin (Ca2+/CaM)-dependent fashion. Ca2+/CaM binding to the N-terminal CaM binding site of GRK5 mediates this effect. Furthermore, GRK5, but not GRK5Δ NLS or GRK2, binds specifically and directly to DNA in vitro. Consistent with their presence in the nuclei of transfected cells, all the GRK4, but not GRK2, subfamily members contain putative NLSs. These results suggest that the GRK4 subfamily of GRKs may play a signaling role in the nucleus and that GRK4 and GRK2 subfamily members perform divergent cellular functions.

CSIG-07. ACTIVATION OF THE ADHESION G PROTEIN-COUPLED RECEPTOR GPR133 BY ANTIBODIES AGAINST THE N-TERMINUS

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi34-vi34
Author(s):  
Gabriele Stephan ◽  
Joshua Frenster ◽  
Niklas Ravn-Boess ◽  
Devin Bready ◽  
Jordan Wilcox ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
Receptor Activation ◽  
Cell Culture Supernatant ◽  
Dependent Manner ◽  
G Protein Coupled Receptor ◽  
Terminal Fragment ◽  
C Terminus ◽  
N Terminus ◽  
G Protein Coupled

Abstract We recently demonstrated that GPR133 (ADGRD1), a member of the adhesion G protein-coupled receptor (aGPCR) family, is necessary for growth of glioblastoma (GBM) and is de novo expressed in GBM relative to normal brain tissue. We therefore postulate that GPR133 represents a novel target in GBM, which merits development of therapeutics. Like most aGPCRs, GPR133 is characterized by an intracellular C-terminus, 7 plasma membrane-spanning α-helices and a large extracellular N-terminus. The N-terminus possesses a conserved GPCR autoproteolysis-inducing (GAIN) domain that catalyzes cleavage at a GPCR proteolysis site (GPS), resulting in a C-terminal fragment (CTF) and an N-terminal fragment (NTF). We showed that dissociation of the cleaved NTF and CTF at the plasma membrane increases canonical signaling of GPR133, which is mediated by coupling to Gs and increase in cytosolic cAMP. Toward characterizing the effect of biologics on GPR133 function, we overexpressed wild-type or mutant forms of GPR133 in HEK293T cells and patient-derived GBM cells lines. Treatment of these cells with antibodies specifically targeting the NTF of GPR133 increased receptor activation in a dose-dependent manner. No effects were elicited with an antibody against the receptor’s intracellular C-terminus. Interestingly, cells overexpressing a cleavage-deficient mutant GPR133 (H543R) did not respond to antibody stimulation, suggesting that the effect is cleavage-dependent. Following antibody treatment, co-purification of the GPR133 NTF and the N-terminal antibody from the cell culture supernatant indicated the formation of antibody-NTF complexes. Analysis of these complexes suggested that antibody binding stimulated the dissociation of the NTF from the CTF. However, the increased flexibility of the GAIN domain and NTF after cleavage, independently of dissociation, may also endow the receptor with responsiveness to the effects of the antibodies. These data constitute a proof-of-concept paradigm of modulation of GPR133 function with antibodies. This work provides rationale for pursuing development of biologics targeting GPR133 in GBM.

scholarly journals Comment on "A G Protein Coupled Receptor Is a Plasma Membrane Receptor for the Plant Hormone Abscisic Acid"

Science ◽  
2007 ◽  
Vol 318 (5852) ◽  
pp. 914c-914c ◽  
Author(s):  
C. A. Johnston ◽  
B. R. Temple ◽  
J.-G. Chen ◽  
Y. Gao ◽  
E. N. Moriyama ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Abscisic Acid ◽  
G Protein ◽  
Plant Hormone ◽  
Membrane Receptor ◽  
G Protein Coupled Receptor ◽  
Plasma Membrane Receptor ◽  
G Protein Coupled

scholarly journals Aromatic D-amino acids act as chemoattractant factors for human leukocytes through a G protein-coupled receptor, GPR109B

2009 ◽  
Vol 106 (10) ◽  
pp. 3930-3934 ◽  
Author(s):  
Y. Irukayama-Tomobe ◽  
H. Tanaka ◽  
T. Yokomizo ◽  
T. Hashidate-Yoshida ◽  
M. Yanagisawa ◽  
...  
Keyword(s):  
Amino Acids ◽  
G Protein ◽  
G Protein Coupled Receptor ◽  
Human Leukocytes ◽  
G Protein Coupled

Cancer driver G-protein coupled receptor (GPCR) induced β-catenin nuclear localization: the transcriptional junction

2017 ◽  
Vol 37 (1) ◽  
pp. 147-157 ◽  
Author(s):  
Jeetendra Kumar Nag ◽  
Tatyana Rudina ◽  
Myriam Maoz ◽  
Sorina Grisaru-Granovsky ◽  
Beatrice Uziely ◽  
...  
Keyword(s):  
G Protein ◽  
Nuclear Localization ◽  
G Protein Coupled Receptor ◽  
Cancer Driver ◽  
G Protein Coupled

scholarly journals Acute depletion of plasma membrane phosphatidylinositol 4,5-bisphosphate impairs specific steps in endocytosis of the G-protein-coupled receptor

2012 ◽  
Vol 125 (9) ◽  
pp. 2185-2197 ◽  
Author(s):  
D. J. Toth ◽  
J. T. Toth ◽  
G. Gulyas ◽  
A. Balla ◽  
T. Balla ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
G Protein ◽  
G Protein Coupled Receptor ◽  
G Protein Coupled
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