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Kinin B1 receptor homo-oligomerization is required for receptor trafficking to the cell surface.

Sandén, Caroline LU and Leeb-Lundberg, Fredrik LU (2012) 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)
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author
organization
publishing date
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
2013-01-02 10:51:26
date last changed
2017-08-13 03:25:24
@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          = {http://dx.doi.org/10.1016/j.intimp.2012.11.012},
  volume       = {15},
  year         = {2012},
}