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Synergistic regulation of beta2-adrenergic receptor sequestration: intracellular complement of beta-adrenergic receptor kinase and beta-arrestin determine kinetics of internalization.

Publication ,  Journal Article
Ménard, L; Ferguson, SS; Zhang, J; Lin, FT; Lefkowitz, RJ; Caron, MG; Barak, LS
Published in: Mol Pharmacol
May 1997

Two of the common mechanisms regulating G protein-coupled receptor (GPCR) signal transduction are phosphorylation and sequestration (internalization). Agonist-mediated receptor phosphorylation by the beta-adrenergic receptor kinase (betaARK) facilitates subsequent interaction with an arrestin protein, resulting in receptor desensitization. Studies of the beta2-adrenergic receptor (beta2AR) receptor in human embryonic kidney (HEK) 293 cells indicate that betaARK and arrestin proteins (beta-arrestins) also regulate sequestration. Consistent with this notion, we show in HEK 293 cells that reduction in or removal of the ability of the beta2AR to be phosphorylated by betaARK or to interact normally with beta-arrestin substantially reduces agonist-mediated sequestration. To evaluate betaARK and beta-arrestin regulation of beta2AR sequestration, we examined the relationship between betaARK and/or beta-arrestin expression and beta2AR sequestration in a variety of cultured cells, including HEK 293, COS 7, CHO, A431, and CHW. COS cells had both the lowest levels of endogenous beta-arrestin expression and beta2AR sequestration, whereas HEK 293 had the highest. Overexpression of beta-arrestin, but not betaARK, in COS cells increased the extent of wild-type beta2AR sequestration to levels observed in HEK 293 cells. However, a betaARK phosphorylation-impaired beta2AR mutant (Y326A) required the simultaneous overexpression of both betaARK and beta-arrestin for this to occur. Among all cell lines, sequestration correlated best with the product of betaARK and beta-arrestin expression. Moreover, an agonist-mediated translocation of wild-type beta2AR and endogenous beta-arrestin 2 to endocytic vesicles prepared from CHO fibroblasts was observed. These data suggest not only that the complement of cellular betaARK and arrestin proteins synergistically regulate beta2AR sequestration but also that beta-arrestins directly regulate beta2AR trafficking as well as desensitization.

Duke Scholars

Published In

Mol Pharmacol

ISSN

0026-895X

Publication Date

May 1997

Volume

51

Issue

5

Start / End Page

800 / 808

Location

United States

Related Subject Headings

  • beta-Arrestins
  • beta-Arrestin 2
  • beta-Adrenergic Receptor Kinases
  • Tumor Cells, Cultured
  • Subcellular Fractions
  • Receptors, Adrenergic, beta-2
  • Phosphorylation
  • Pharmacology & Pharmacy
  • Kinetics
  • Humans
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Ménard, L., Ferguson, S. S., Zhang, J., Lin, F. T., Lefkowitz, R. J., Caron, M. G., & Barak, L. S. (1997). Synergistic regulation of beta2-adrenergic receptor sequestration: intracellular complement of beta-adrenergic receptor kinase and beta-arrestin determine kinetics of internalization. Mol Pharmacol, 51(5), 800–808.
Ménard, L., S. S. Ferguson, J. Zhang, F. T. Lin, R. J. Lefkowitz, M. G. Caron, and L. S. Barak. “Synergistic regulation of beta2-adrenergic receptor sequestration: intracellular complement of beta-adrenergic receptor kinase and beta-arrestin determine kinetics of internalization.Mol Pharmacol 51, no. 5 (May 1997): 800–808.

Published In

Mol Pharmacol

ISSN

0026-895X

Publication Date

May 1997

Volume

51

Issue

5

Start / End Page

800 / 808

Location

United States

Related Subject Headings

  • beta-Arrestins
  • beta-Arrestin 2
  • beta-Adrenergic Receptor Kinases
  • Tumor Cells, Cultured
  • Subcellular Fractions
  • Receptors, Adrenergic, beta-2
  • Phosphorylation
  • Pharmacology & Pharmacy
  • Kinetics
  • Humans