N‐glycosylation of the β<sub>2</sub> adrenergic receptor regulates receptor function by modulating dimerization

Li, Xiaona; Zhou, Mang; Huang, Wei; Yang, Huaiyu

doi:10.1111/febs.14098

Cited by 14 publications

(11 citation statements)

References 49 publications

(74 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the reason for this loss of beta adrenergic receptor activity is not fully understood, differences observed in the expression of β1and β3-AR may provide at least a partial explanation. In addition, our pharmacological data showed that β3AR was almost entirely glycosylated in M+T1AM cells, suggesting that β3-AR might be desensitized, as this post-translational modification has been known to play an important role in receptor function [26] and indicated, for the first time, the potentiality of T1AM to affect protein function by promoting post-translational modifications. This finding indicated that T1AM treatment might lead to constitutive activation of β3AR, leading to desensitization of this receptor.…”

Section: Discussionmentioning

confidence: 68%

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

Gencarelli

Laurino

Landucci

et al. 2020

Biology

View full text Add to dashboard Cite

We investigated the effect of 3-iodothyronamine (T1AM) on thermogenic substrates in brown adipocytes (BAs). BAs isolated from the stromal fraction of rat brown adipose tissue were exposed to an adipogenic medium containing insulin in the absence (M) or in the presence of 20 nM T1AM (M+T1AM) for 6 days. At the end of the treatment, the expression of p-PKA/PKA, p-AKT/AKT, p-AMPK/AMPK, p-CREB/CREB, p-P38/P38, type 1 and 3 beta adrenergic receptors (β1–β3AR), GLUT4, type 2 deiodinase (DIO2), and uncoupling protein 1 (UCP-1) were evaluated. The effects of cell conditioning with T1AM on fatty acid mobilization (basal and adrenergic-mediated), glucose uptake (basal and insulin-mediated), and ATP cell content were also analyzed in both cell populations. When compared to cells not exposed, M+T1AM cells showed increased p-PKA/PKA, p-AKT/AKT, p-CREB/CREB, p-P38/P38, and p-AMPK/AMPK, downregulation of DIO2 and β1AR, and upregulation of glycosylated β3AR, GLUT4, and adiponectin. At basal conditions, glycerol release was higher for M+T1AM cells than M cells, without any significant differences in basal glucose uptake. Notably, in M+T1AM cells, adrenergic agonists failed to activate PKA and lipolysis and to increase ATP level, but the glucose uptake in response to insulin exposure was more pronounced than in M cells. In conclusion, our results suggest that BAs conditioning with T1AM promote a catabolic condition promising to fight obesity and insulin resistance.

show abstract

Section: Discussionmentioning

confidence: 68%

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

Gencarelli

Laurino

Landucci

et al. 2020

Biology

View full text Add to dashboard Cite

show abstract

“…Previous studies have shown that β2‐adrenergic receptor can form SDS‐resistant (Hebert et al., 1996) and SDS‐sensitive homodimers (Angers et al., 2000). Recently, N‐linked glycosylation of the extracellular N‐terminal domain has been found to be involved in receptor dimerization of the human bradykinin B2 (Michineau et al., 2006) and β2‐adrenergic receptors (Li et al., 2017). In this study, in SDS‐PAGE experiments with anti‐Flag antibody, two bands, 45 and 85 kDa corresponding to di‐ and mono‐species, were detected in both HEK‐293T and DU145 cells.…”

Section: Discussionmentioning

confidence: 99%

N‐glycosylation of the human neuropeptide QRFP receptor (QRFPR) is essential for ligand binding and receptor activation

Wang

Yan

Shi

et al. 2021

Journal of Neurochemistry

View full text Add to dashboard Cite

The newly identified pyroglutamylated RFamide peptide (QRFP) signaling system has been shown to be implicated in regulating a variety of physiological processes. G-protein-coupled receptors (GPCRs) are preferentially N-glycosylated on extracellular domains. The human QRFP receptor QRFPR (GPR103) possesses three Nglycosylation consensus sites, two located on the N-terminal domain (N5 and N19) and one on the first extracellular loop (ECL1) (N106); however, to date, their role in QRFPR expression and signaling has not been established. Here, we combined mutants with glutamine substitution of the critical asparagines of the consensus sites with glycosidase PNGase F and N-glycosylation inhibitor tunicamycin to study the effect of N-glycosylation in the regulation of QRFPR cell surface expression and signaling. Western blot analysis performed with site-directed mutagenesis revealed that two asparagines at N19 in the N-terminus and N106 in ECL1, but not N5 in the N-terminus, served as sites for N-glycosylation. Treatment with PNGase F and tunicamycin resulted in a reduction in both two-protein species, ~43 kDa and ~85 kDa in size, by 2-4 kDa. Analysis with confocal microscopy and quantitative ELISA showed that N-glycosylation of QRFPR is not essentially required for targeting the cell membrane. However, further binding assay and functional assays demonstrated that removal of N-glycosylation sequons or treatment with tunicamycin led to significant impairments in the interaction of receptor with QRFP26 and downstream signaling. Thus, our findings suggest that for the human QRFP receptor (QRFPR), N-glycosylation is not important for cell surface expression but is a pre-requisite for ligand binding and receptor activation.

show abstract

“…Since N-linked glycosylation is a critical step in the maturation of RTKs, its disruption has been proposed to be an effective strategy to target both primary and redundant RTK signaling 24,28 . However, normal cells may also rely on this pathway for their growth and development 29,30 . Nevertheless, suppression of GluIIβ may represent a novel approach to selectively inhibit N-linked glycosylation and thus specifically reduce RTKs activities in tumor cells.…”

Section: Discussionmentioning

confidence: 99%

Knockout of glucosidase II beta subunit inhibits growth and metastatic potential of lung cancer cells by inhibiting receptor tyrosine kinase activities

Khaodee

Udomsom

Kunnaja

et al. 2019

Sci Rep

View full text Add to dashboard Cite

Glucosidase II (GluII) plays a major role in regulating post-translation modification of N-linked glycoproteins. We have previously reported that the expression of glucosidase II beta subunit (GluIIβ) was significantly increased in lung tumor tissues and its suppression triggers autophagy and/or apoptosis. Here, we investigated the role of GluIIβ in cell growth, metastatic potential, and receptor tyrosine kinases (RTKs) signaling activity in lung carcinoma cell lines. CRISPR-CAS9 technology was used to knockout the GluIIβ encoding gene (PRKSH) in lung carcinoma cells. GluIIβ knockout cells exhibited drastically slower growth rates in comparison to non-target transfected cells, particularly with lower concentrations of fetal bovine serum, indicating impairment of their ability to survive under nutritional deprivation. Cell migration and anchorage-independent growth, the fundamental components of cancer cell metastasis, were significantly decreased in GluIIβ knockout cells. Knockout of GluIIβ increased the sensitivity of lung cancer cells to cisplatin but reduced their sensitivity to gefitinib. Interestingly, knocking out of GluIIβ lowered overall RTK signaling activities to less than half of those in non-target transfected cells, which could represent a novel strategy for blocking multiple RTKs in tumor cells in an effort to improve lung cancer treatment.

show abstract

N‐glycosylation of the β₂ adrenergic receptor regulates receptor function by modulating dimerization

Abstract: Peptide-N-glycosidase F (PNGase F, EC 3.2.2.11); endo-β-N-acetylglucosaminidase A (Endo-A, EC 3.2.1.96).

Cited by 14 publications

References 49 publications

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

N‐glycosylation of the human neuropeptide QRFP receptor (QRFPR) is essential for ligand binding and receptor activation

Knockout of glucosidase II beta subunit inhibits growth and metastatic potential of lung cancer cells by inhibiting receptor tyrosine kinase activities

Contact Info

Product

Resources

About

N‐glycosylation of the β2 adrenergic receptor regulates receptor function by modulating dimerization

Abstract: Peptide-N-glycosidase F (PNGase F, EC 3.2.2.11); endo-β-N-acetylglucosaminidase A (Endo-A, EC 3.2.1.96).

Cited by 14 publications

References 49 publications

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

3-Iodothyronamine Affects Thermogenic Substrates’ Mobilization in Brown Adipocytes

N‐glycosylation of the human neuropeptide QRFP receptor (QRFPR) is essential for ligand binding and receptor activation

Knockout of glucosidase II beta subunit inhibits growth and metastatic potential of lung cancer cells by inhibiting receptor tyrosine kinase activities

Contact Info

Product

Resources

About

N‐glycosylation of the β₂ adrenergic receptor regulates receptor function by modulating dimerization