Advanced

Human B1 and B2 bradykinin receptors and their agonists target caveolae-related lipid rafts to different degrees in HEK293 cells

Lamb, ME; Zhang, CW; Shea, T; Kyle, DJ and Leeb-Lundberg, Fredrik LU (2002) In Biochemistry 41(48). p.14340-14347
Abstract
To address the targeting of G protein-coupled receptors to caveolae-related lipid rafts (CLR), we studied the human B2 (B2R) and B1 (B1R) bradykinin receptor subtypes in HEK293 cells. CLR were enriched on the basis of their unique buoyant density and composition of cholesterol, caveolin-1, and flotillin-1 but not clathrin. CLR contained B2R and B1R as determined by both receptor immunoblotting and the increase in specific activity of receptor agonist binding to cells at both 4 and 37 degreesC when binding was followed by CLR enrichment. B2R was highly enriched in this fraction, whereas B I R was not enriched. Furthermore, acid washing of cells prior to cell disruption minimally affected the CLR-associated B2R agonist binding, whereas it... (More)
To address the targeting of G protein-coupled receptors to caveolae-related lipid rafts (CLR), we studied the human B2 (B2R) and B1 (B1R) bradykinin receptor subtypes in HEK293 cells. CLR were enriched on the basis of their unique buoyant density and composition of cholesterol, caveolin-1, and flotillin-1 but not clathrin. CLR contained B2R and B1R as determined by both receptor immunoblotting and the increase in specific activity of receptor agonist binding to cells at both 4 and 37 degreesC when binding was followed by CLR enrichment. B2R was highly enriched in this fraction, whereas B I R was not enriched. Furthermore, acid washing of cells prior to cell disruption minimally affected the CLR-associated B2R agonist binding, whereas it dissociated a major portion of the CLR-associated B I R agonist binding. In addition, when agonist binding at 4 degreesC was followed by an increase in the temperature to 37 degreesC B2R agonist binding in CLR transiently increased, and this increase was dependent on the C-terminal domain. On the other hand, B1R agonist binding remained unchanged and was independent of the C-terminal domain. Our results show that B2R is constitutively targeted to CLR in HEK293 cells and appears to shuttle the agonist through these domains, whereas B1R may be there by default. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biochemistry
volume
41
issue
48
pages
14340 - 14347
publisher
The American Chemical Society
external identifiers
  • wos:000179517000030
  • scopus:2242478438
ISSN
0006-2960
DOI
10.1021/bi020231d
language
English
LU publication?
yes
id
99ad45a3-bb4f-4717-84fe-c41802db9d9e (old id 322692)
date added to LUP
2007-11-05 15:55:53
date last changed
2017-05-28 03:45:46
@article{99ad45a3-bb4f-4717-84fe-c41802db9d9e,
  abstract     = {To address the targeting of G protein-coupled receptors to caveolae-related lipid rafts (CLR), we studied the human B2 (B2R) and B1 (B1R) bradykinin receptor subtypes in HEK293 cells. CLR were enriched on the basis of their unique buoyant density and composition of cholesterol, caveolin-1, and flotillin-1 but not clathrin. CLR contained B2R and B1R as determined by both receptor immunoblotting and the increase in specific activity of receptor agonist binding to cells at both 4 and 37 degreesC when binding was followed by CLR enrichment. B2R was highly enriched in this fraction, whereas B I R was not enriched. Furthermore, acid washing of cells prior to cell disruption minimally affected the CLR-associated B2R agonist binding, whereas it dissociated a major portion of the CLR-associated B I R agonist binding. In addition, when agonist binding at 4 degreesC was followed by an increase in the temperature to 37 degreesC B2R agonist binding in CLR transiently increased, and this increase was dependent on the C-terminal domain. On the other hand, B1R agonist binding remained unchanged and was independent of the C-terminal domain. Our results show that B2R is constitutively targeted to CLR in HEK293 cells and appears to shuttle the agonist through these domains, whereas B1R may be there by default.},
  author       = {Lamb, ME and Zhang, CW and Shea, T and Kyle, DJ and Leeb-Lundberg, Fredrik},
  issn         = {0006-2960},
  language     = {eng},
  number       = {48},
  pages        = {14340--14347},
  publisher    = {The American Chemical Society},
  series       = {Biochemistry},
  title        = {Human B1 and B2 bradykinin receptors and their agonists target caveolae-related lipid rafts to different degrees in HEK293 cells},
  url          = {http://dx.doi.org/10.1021/bi020231d},
  volume       = {41},
  year         = {2002},
}