Interhelical Interaction and Receptor Phosphorylation Regulate the Activation Kinetics of Different Human β1-Adrenoceptor Variants

Ahles, Andrea; Rodewald, Fabian; Rochais, Francesca; Bünemann, Moritz; Engelhardt, Stefan

doi:10.1074/jbc.m114.607333

Cited by 11 publications

(9 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With a FRET-based biosensor for ␤ 1 -AR termed ␤ 1 -AR sensor (20) we directly measured depolarization-mediated inactivating conformational changes occurring with very fast kinetics. FRET assays between ␤ 1 -AR and either a G protein subunit (21) or arrestin 3 (Arr3) (22,23) showed deactivation effects of depolarization on downstream signaling. The closely related ␤ 2 -AR was also voltage-dependent but the extent of depolarization-induced deactivation was less pronounced than in ␤ 1 -AR.…”

mentioning

confidence: 99%

Membrane Potential Controls the Efficacy of Catecholamine-induced β1-Adrenoceptor Activity

Birk

Rinne

Bünemann

2015

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Background:The activity of several G q -and G i -coupled receptors is modulated by the membrane potential. Results: Voltage modulates catecholamine-mediated activation of G s -coupled ␤ 1 -and ␤ 2 -adrenoceptors. Conclusion: Voltage-dependence of ␤ 1 -AR is due to alterations in the efficacy of catecholamines. Significance: By modulating catecholamine efficacy on ␤ 1 -ARs, voltage can modify receptor activity on a very fast time scale.

show abstract

mentioning

confidence: 99%

Membrane Potential Controls the Efficacy of Catecholamine-induced β1-Adrenoceptor Activity

Birk

Rinne

Bünemann

2015

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…In most GPCRs, genetic variation within GPCRs leads to their cellular signaling transduction, phenotypes, and variable drug responses. 5 , 30 , 31 , 32 To date, numerous non-synonymous genetic variations have been identified in humans. SNPs can be assigned a MAF, which is the less common allele frequency at a locus that is observed in a population.…”

Section: Discussionmentioning

confidence: 99%

Leukotriene B4 receptor 2 gene polymorphism (rs1950504, Asp196Gly) leads to enhanced cell motility under low-dose ligand stimulation

et al. 2017

View full text Add to dashboard Cite

Recently, single-nucleotide polymorphisms (SNPs) in G-protein-coupled receptors (GPCRs) have been suggested to contribute to physiopathology and therapeutic effects. Leukotriene B4 receptor 2 (BLT2), a member of the GPCR family, plays a critical role in the pathogenesis of several inflammatory diseases, including cancer and asthma. However, no studies on BLT2 SNP effects have been reported to date. In this study, we demonstrate that the BLT2 SNP (rs1950504, Asp196Gly), a Gly-196 variant of BLT2 (BLT2 D196G), causes enhanced cell motility under low-dose stimulation of its ligands. In addition, we demonstrated that Akt activation and subsequent production of reactive oxygen species (ROS), both of which act downstream of BLT2, are also increased by BLT2 D196G in response to low-dose ligand stimulation. Furthermore, we observed that the ligand binding affinity of BLT2 D196G was enhanced compared with that of BLT2. Through homology modeling analysis, it was predicted that BLT2 D196G loses ionic interaction with R197, potentially resulting in increased agonist-receptor interaction. To the best of our knowledge, this report is the first to describe a SNP study on BLT2 and shows that BLT2 D196G enhances ligand sensitivity, thereby increasing cell motility in response to low-dose ligand stimulation.

show abstract

“…The different ADRB1 mutants for the third intracellular loop and the C-terminus were generated according to the presence of restriction enzyme binding sites. YFP-β-arrestin2 was generated as described previously [ 26 ]. Cer was fused to the β 1 -adrenoceptor’s C-terminus with a linker of five glycine residues.…”

Section: Methodsmentioning

confidence: 99%

“…Radioactive phosphorylation assays were performed as described previously [ 26 ] with the exception that cells were grown in DMEM containing 1% FCS and 1% penicillin/ streptomycin. As a loading control the same membranes were used to perform a western blot analysis detecting total ADRB1.…”

Section: Methodsmentioning

confidence: 99%

Two serines in the distal C-terminus of the human ß1-adrenoceptor determine ß-arrestin2 recruitment

et al. 2017

Self Cite

View full text Add to dashboard Cite

G protein-coupled receptors (GPCRs) undergo phosphorylation at several intracellular residues by G protein-coupled receptor kinases. The resulting phosphorylation pattern triggers arrestin recruitment and receptor desensitization. The exact sites of phosphorylation and their function remained largely unknown for the human β1-adrenoceptor (ADRB1), a key GPCR in adrenergic signal transduction and the target of widely used drugs such as β-blockers. The present study aimed to identify the intracellular phosphorylation sites in the ADRB1 and to delineate their function. The human ADRB1 was expressed in HEK293 cells and its phosphorylation pattern was determined by mass spectrometric analysis before and after stimulation with a receptor agonist. We identified a total of eight phosphorylation sites in the receptor’s third intracellular loop and C-terminus. Analyzing the functional relevance of individual sites using phosphosite-deficient receptor mutants we found phosphorylation of the ADRB1 at Ser461/Ser462 in the distal part of the C-terminus to determine β-arrestin2 recruitment and receptor internalization. Our data reveal the phosphorylation pattern of the human ADRB1 and the site that mediates recruitment of β-arrestin2.

show abstract

Interhelical Interaction and Receptor Phosphorylation Regulate the Activation Kinetics of Different Human β1-Adrenoceptor Variants

Cited by 11 publications

References 34 publications

Membrane Potential Controls the Efficacy of Catecholamine-induced β1-Adrenoceptor Activity

Membrane Potential Controls the Efficacy of Catecholamine-induced β1-Adrenoceptor Activity

Leukotriene B4 receptor 2 gene polymorphism (rs1950504, Asp196Gly) leads to enhanced cell motility under low-dose ligand stimulation

Two serines in the distal C-terminus of the human ß1-adrenoceptor determine ß-arrestin2 recruitment

Contact Info

Product

Resources

About