Kinin B1 receptor homo-oligomerization is required for receptor trafficking to the cell surface.
(2013) In International Immunopharmacology 15(1). p.121-128- Abstract
- The kinin B1 receptor (B1R) is a G protein-coupled receptor with pro-inflammatory activity that is latent in healthy tissues but induced by tissue insult. Here, we investigated if B1R homo-oligomerization is a possible mechanism regulating the presentation of this receptor at the level of maturation and trafficking to the cell surface. To this end, we used HEK293 cells stably expressing N-terminal FLAG and HA epitope-tagged wild-type human B1R and an N-terminal receptor fragment, B1stop135, which terminates at the C-terminal end of the third transmembrane domain and has previously been shown to oligomerize with B1R. Receptors were monitored by immunoblotting and immunoprecipitation, receptor function by agonist binding and agonist-promoted... (More)
- The kinin B1 receptor (B1R) is a G protein-coupled receptor with pro-inflammatory activity that is latent in healthy tissues but induced by tissue insult. Here, we investigated if B1R homo-oligomerization is a possible mechanism regulating the presentation of this receptor at the level of maturation and trafficking to the cell surface. To this end, we used HEK293 cells stably expressing N-terminal FLAG and HA epitope-tagged wild-type human B1R and an N-terminal receptor fragment, B1stop135, which terminates at the C-terminal end of the third transmembrane domain and has previously been shown to oligomerize with B1R. Receptors were monitored by immunoblotting and immunoprecipitation, receptor function by agonist binding and agonist-promoted phosphoinositide hydrolysis, and receptor trafficking by confocal immunofluorescence microscopy. When expressed alone, B1R is core N-glycosylated and forms oligomers localized intracellularly and on the cell surface. B1stop135 also exists as core N-glycosylated oligomers but is localized exclusively intracellularly. When co-expressed, B1stop135 prevents specifically B1R homo-oligomerization by forming nonfunctional B1R-B1stop135 hetero-oligomers, retains B1R intracellularly at least in part in the endoplasmatic reticulum (ER), increases calnexin binding to the receptor, and increases receptor degradation. We conclude that B1R homo-oligomerization is necessary for B1R maturation and trafficking to the cell surface. Modulating this mechanism may be a novel therapeutic avenue in inflammatory disease. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3347785
- author
- Sandén, Caroline LU and Leeb-Lundberg, Fredrik LU
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- in
- International Immunopharmacology
- volume
- 15
- issue
- 1
- pages
- 121 - 128
- publisher
- Elsevier
- external identifiers
-
- wos:000315075400017
- pmid:23201435
- scopus:84872350044
- ISSN
- 1878-1705
- DOI
- 10.1016/j.intimp.2012.11.012
- language
- English
- LU publication?
- yes
- id
- ed7e9cb0-1443-4436-9597-54b2590edf9f (old id 3347785)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/23201435?dopt=Abstract
- date added to LUP
- 2016-04-01 11:11:27
- date last changed
- 2022-01-26 06:07:16
@article{ed7e9cb0-1443-4436-9597-54b2590edf9f,
abstract = {{The kinin B1 receptor (B1R) is a G protein-coupled receptor with pro-inflammatory activity that is latent in healthy tissues but induced by tissue insult. Here, we investigated if B1R homo-oligomerization is a possible mechanism regulating the presentation of this receptor at the level of maturation and trafficking to the cell surface. To this end, we used HEK293 cells stably expressing N-terminal FLAG and HA epitope-tagged wild-type human B1R and an N-terminal receptor fragment, B1stop135, which terminates at the C-terminal end of the third transmembrane domain and has previously been shown to oligomerize with B1R. Receptors were monitored by immunoblotting and immunoprecipitation, receptor function by agonist binding and agonist-promoted phosphoinositide hydrolysis, and receptor trafficking by confocal immunofluorescence microscopy. When expressed alone, B1R is core N-glycosylated and forms oligomers localized intracellularly and on the cell surface. B1stop135 also exists as core N-glycosylated oligomers but is localized exclusively intracellularly. When co-expressed, B1stop135 prevents specifically B1R homo-oligomerization by forming nonfunctional B1R-B1stop135 hetero-oligomers, retains B1R intracellularly at least in part in the endoplasmatic reticulum (ER), increases calnexin binding to the receptor, and increases receptor degradation. We conclude that B1R homo-oligomerization is necessary for B1R maturation and trafficking to the cell surface. Modulating this mechanism may be a novel therapeutic avenue in inflammatory disease.}},
author = {{Sandén, Caroline and Leeb-Lundberg, Fredrik}},
issn = {{1878-1705}},
language = {{eng}},
number = {{1}},
pages = {{121--128}},
publisher = {{Elsevier}},
series = {{International Immunopharmacology}},
title = {{Kinin B1 receptor homo-oligomerization is required for receptor trafficking to the cell surface.}},
url = {{https://lup.lub.lu.se/search/files/2454361/3632121.pdf}},
doi = {{10.1016/j.intimp.2012.11.012}},
volume = {{15}},
year = {{2013}},
}