B1 Bradykinin Receptor Homo-oligomers in Receptor Cell Surface Expression and Signaling: Effects of Receptor Fragments.
(2005) In Molecular Pharmacology 67(1). p.309-318- Abstract
- The human B1 bradykinin receptor is an inducible and constitutively active G protein-coupled receptor that is involved in the inflammatory and pain responses to injury. Here, we investigated the role of B1 receptor homo-oligomerization in cell surface receptor expression. B1 receptors tagged with either the FLAG or hemagglutinin epitope were monitored immunologically and by radio-ligand binding, biotinylation, and phosphoinositide hydrolysis in human embryonic kidney 293 cells. Selective immunoprecipitation, immunoblotting, and immunoelectron microscopy with epitope-specific antibodies together provided evidence for constitutively formed cell surface receptor homo-oligomers. Truncation of the receptor from the N- and C-terminal ends... (More)
- The human B1 bradykinin receptor is an inducible and constitutively active G protein-coupled receptor that is involved in the inflammatory and pain responses to injury. Here, we investigated the role of B1 receptor homo-oligomerization in cell surface receptor expression. B1 receptors tagged with either the FLAG or hemagglutinin epitope were monitored immunologically and by radio-ligand binding, biotinylation, and phosphoinositide hydrolysis in human embryonic kidney 293 cells. Selective immunoprecipitation, immunoblotting, and immunoelectron microscopy with epitope-specific antibodies together provided evidence for constitutively formed cell surface receptor homo-oligomers. Truncation of the receptor from the N- and C-terminal ends indicated that the epitope for oligomerization seems to be located between Leu26 on top of transmembrane helix 1 and Val71 at the bottom of helix 2. A receptor construct terminating at Asp134 at the bottom of helix 3, B1stop135, was expressed in the cell. It is interesting that this construct behaved as a dominant-negative mutant by competitively preventing formation of intact B1 receptor homo-oligomers, and redistributing B1 receptors from the cell surface to a common intracellular compartment. In contrast, expression of a construct containing the residues downstream of Asp134, B1del(2-134), was inactive in this regard. Together, these results are consistent with a mechanism where constitutive B1 receptor homooligomerization is required for expression of receptors on the cell surface and subsequent constitutive receptor signaling. This may be a novel mechanism by which the cell regulates the presentation of this constitutively highly active receptor at various stages of injury. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/129779
- author
- Kang, Dongsoo LU ; Gustafsson, Caroline ; Mörgelin, Matthias LU and Leeb-Lundberg, Fredrik LU
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Molecular Pharmacology
- volume
- 67
- issue
- 1
- pages
- 309 - 318
- publisher
- American Society for Pharmacology and Experimental Therapeutics
- external identifiers
-
- pmid:15492119
- wos:000225865200035
- scopus:11244319585
- ISSN
- 1521-0111
- DOI
- 10.1124/mol.104.002840
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Department of Experimental Medical Science (013210000), Molecular Neurobiology (0131000140), Drug Target Discovery (013212045), Division of Infection Medicine (BMC) (013024020)
- id
- e89a0038-923c-49fc-b45f-4806e76022e9 (old id 129779)
- date added to LUP
- 2016-04-01 12:32:20
- date last changed
- 2022-04-21 08:49:27
@article{e89a0038-923c-49fc-b45f-4806e76022e9,
abstract = {{The human B1 bradykinin receptor is an inducible and constitutively active G protein-coupled receptor that is involved in the inflammatory and pain responses to injury. Here, we investigated the role of B1 receptor homo-oligomerization in cell surface receptor expression. B1 receptors tagged with either the FLAG or hemagglutinin epitope were monitored immunologically and by radio-ligand binding, biotinylation, and phosphoinositide hydrolysis in human embryonic kidney 293 cells. Selective immunoprecipitation, immunoblotting, and immunoelectron microscopy with epitope-specific antibodies together provided evidence for constitutively formed cell surface receptor homo-oligomers. Truncation of the receptor from the N- and C-terminal ends indicated that the epitope for oligomerization seems to be located between Leu26 on top of transmembrane helix 1 and Val71 at the bottom of helix 2. A receptor construct terminating at Asp134 at the bottom of helix 3, B1stop135, was expressed in the cell. It is interesting that this construct behaved as a dominant-negative mutant by competitively preventing formation of intact B1 receptor homo-oligomers, and redistributing B1 receptors from the cell surface to a common intracellular compartment. In contrast, expression of a construct containing the residues downstream of Asp134, B1del(2-134), was inactive in this regard. Together, these results are consistent with a mechanism where constitutive B1 receptor homooligomerization is required for expression of receptors on the cell surface and subsequent constitutive receptor signaling. This may be a novel mechanism by which the cell regulates the presentation of this constitutively highly active receptor at various stages of injury.}},
author = {{Kang, Dongsoo and Gustafsson, Caroline and Mörgelin, Matthias and Leeb-Lundberg, Fredrik}},
issn = {{1521-0111}},
language = {{eng}},
number = {{1}},
pages = {{309--318}},
publisher = {{American Society for Pharmacology and Experimental Therapeutics}},
series = {{Molecular Pharmacology}},
title = {{B1 Bradykinin Receptor Homo-oligomers in Receptor Cell Surface Expression and Signaling: Effects of Receptor Fragments.}},
url = {{http://dx.doi.org/10.1124/mol.104.002840}},
doi = {{10.1124/mol.104.002840}},
volume = {{67}},
year = {{2005}},
}