Differences in endosomal targeting of human β1- and β2-adrenergic receptors following clathrin-mediated endocytosis

Liang, Wei; Curran, Patricia K.; Hoang, Quang; Moreland, Russell; Fishman, Peter H.

doi:10.1242/jcs.00878

Cited by 33 publications

(45 citation statements)

References 54 publications

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“…Rabbit antihuman ␤ 1 AR (A-20) and ␤ 2 AR (H-20) and mouse monoclonal antiubiquitin IgG 1 Ub(P4D1) were from Santa Cruz Biotechnology, Alexa Fluor 488-conjugated mouse anti-HA.11 was from Covance; mouse monoclonal anti-human LAMP1 IgG 1 (H4A3) was from BD Biosciences, and Cy3-conjugated donkey anti-mouse was from Jackson ImmunoResearch. Sources of other reagents were described previously (27)(28)(29). Plasmids pcDNA3.1-h␤ 1 AR, -h␤ 2 AR, -HA-h␤ 1 AR, -HA-h␤ 1 /␤ 2 ct-AR, and -HA-h␤ 2 /␤ 1 ct-AR, pE-h␤ 1 AR-GFP, and pcDNA3-h␤ 2 AR-GFP, -arrestin-2 and -3 were described before (27,29).…”

Section: Methodsmentioning

confidence: 99%

“…Sources of other reagents were described previously (27)(28)(29). Plasmids pcDNA3.1-h␤ 1 AR, -h␤ 2 AR, -HA-h␤ 1 AR, -HA-h␤ 1 /␤ 2 ct-AR, and -HA-h␤ 2 /␤ 1 ct-AR, pE-h␤ 1 AR-GFP, and pcDNA3-h␤ 2 AR-GFP, -arrestin-2 and -3 were described before (27,29).…”

Section: Methodsmentioning

confidence: 99%

“…Clonal lines stably expressing either ␤ 1 AR or ␤ 2 AR or co-expressing both arrestin-2 and either subtype were described previously (27,29). To obtain cells expressing wild type or chimeric HA-␤AR or ␤AR-GFP or co-expressing one of these receptors and either arrestin-2 or -3, transfections were done with the appropriate plasmid(s) using LipofectAMINE Plus.…”

Section: Methodsmentioning

confidence: 99%

“…Because ␤ 1 AR is associated with human diseases such as congestive heart failure (18) and depression (20), the regulation of ␤ 1 AR is of considerable interest. In comparison to the human ␤ 2 AR, the human ␤ 1 AR is more resistant to agonist-mediated desensitization (21,22), internalization (21,(23)(24)(25)(26)(27), and downregulation (21,23,26,28,29). We have previously shown that in agonist-stimulated BHK cells, ␤ 1 AR is up-regulated due to its resistance to degradation and its increased synthesis (29).…”

mentioning

confidence: 99%

“…We have previously shown that in agonist-stimulated BHK cells, ␤ 1 AR is up-regulated due to its resistance to degradation and its increased synthesis (29). We recently demonstrated that ␤ 1 AR expressed in BHK and HEK 293 cells undergoes agonist-mediated endocytosis by the clathrin-coated pit pathway but traffics to an endosomal compartment distinct from that of ␤ 2 AR (27). Thus, differences in subtype degradation may be due to differences in subtype trafficking.…”

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confidence: 99%

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Resistance of the Human β1-Adrenergic Receptor to Agonist-induced Ubiquitination

Liang¹,

Fishman²

2004

Journal of Biological Chemistry

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Down-regulation is a classic response of most G protein-coupled receptors to prolonged agonist stimulation. We recently showed that when expressed in baby hamster kidney cells, the human ␤ 1 -but not the ␤ 2 -adrenergic receptor (AR) is totally resistant to agonistmediated down-regulation, whereas both have similar rates of basal degradation (Liang, W., Austin, S., Hoang, Q., and Fishman, P. H. (2003) J. Biol. Chem. 278, 39773-39781). To identify the underlying mechanism(s) for this resistance, we investigated the role of proteasomes, lysosomes, and ubiquitination in the degradation of ␤ 1 AR expressed in baby hamster kidney and human embryonic kidney 293 cells. Both lysosomal and proteasomal inhibitors reduced ␤ 1 AR degradation in agonist-stimulated cells but were less effective on basal degradation. To determine whether ␤ 1 AR trafficked to lysosomes we used confocal fluorescence microscopy. We observed some colocalization of ␤ 1 AR and lysosomal markers in agonist-treated cells but much less than that of ␤ 2 AR even in cells co-transfected with arrestin-2, which increases ␤ 1 AR internalization. Ubiquitination of ␤ 2 AR readily occurred in agonist-stimulated cells, whereas ubiquitination of ␤ 1 AR was not detectable even under conditions optimal for that of ␤ 2 AR. Moreover, in cells expressing ␤AR chimeras in which the C termini have been switched, the chimeric ␤ 1 AR with ␤ 2 AR C-tail underwent ubiquitination and down-regulation, but the chimeric ␤ 2 AR with ␤ 1 AR C-tail did not. Our results demonstrate for the first time that ␤ 1 AR and ␤ 2 AR differ in the ability to be ubiquitinated. Because ubiquitin serves as a signal for sorting membrane receptors to lysosomes, the lack of agonist-mediated ubiquitination of ␤ 1 AR may prevent its extensive trafficking to lysosomes and, thus, account for its resistance to down-regulation.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Resistance of the Human β1-Adrenergic Receptor to Agonist-induced Ubiquitination

Liang¹,

Fishman²

2004

Journal of Biological Chemistry

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β1-Adrenergic Receptor Recycles Via a Membranous Organelle, Recycling Endosome, by Binding with Sorting Nexin27

Nakagawa

Asahi

2013

J Membrane Biol

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In cardiomyocytes, β1-adrenergic receptor (β1-AR) plays an important role in regulating cardiac functions. Upon continuous ligand stimulation, β1-AR is internalized and mostly recycled back to the plasma membrane (PM). The recycling endosome (RE) is one of the membranous organelles involved in the protein recycling pathway. To determine whether RE is involved in the internalization of β1-AR upon ligand stimulation, we evaluated the localization of β1-AR after stimulation with a β-agonist, isoproterenol (Iso), in β1-AR-transfected COS-1 cells. After 30 min of Iso treatment and cell surface labeling with the appropriate antibodies, β1-AR was internalized from PM and translocated into the perinuclear region, the same location as the transferrin receptor, an RE marker. We then evaluated whether sorting nexin 27 (SNX27) participated in the β1-AR recycling pathway. When β1-AR and SNX27 were coexpressed, β1-AR coimmunoprecipitated with SNX27. In addition, shRNA-mediated silencing of SNX27 compromised β1-AR recycling and enhanced its delivery into lysosome. Overall, β1-AR on PM was internalized into RE upon Iso stimulation and recycled by RE through binding with SNX27 in COS-1 cells.

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Cell therapy rescues aging-induced beta-1 adrenergic receptor and GRK2 dysfunction in the coronary microcirculation

et al. 2021

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Differences in endosomal targeting of human β1- and β2-adrenergic receptors following clathrin-mediated endocytosis

Abstract: SummaryDifferences in endosomal targeting of human β β 1 -and β β 2 -adrenergic receptors following clathrin-mediated endocytosis

Cited by 33 publications

References 54 publications

Resistance of the Human β1-Adrenergic Receptor to Agonist-induced Ubiquitination

Resistance of the Human β1-Adrenergic Receptor to Agonist-induced Ubiquitination

β1-Adrenergic Receptor Recycles Via a Membranous Organelle, Recycling Endosome, by Binding with Sorting Nexin27

Cell therapy rescues aging-induced beta-1 adrenergic receptor and GRK2 dysfunction in the coronary microcirculation

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