Receptor/beta-arrestin complex formation and the differential trafficking and resensitization of beta2-adrenergic and angiotensin II type 1A receptors

Mol Endocrinol. 2000 Dec;14(12):2040-53. doi: 10.1210/mend.14.12.0565.

Abstract

Beta-arrestins target G protein-coupled receptors (GPCRs) for endocytosis via clathrin-coated vesicles. Beta-arrestins also become detectable on endocytic vesicles in response to angiotensin II type 1A receptor (AT1AR), but not beta2-adrenergic receptor (beta2AR), activation. The carboxyl-terminal tails of these receptors contribute directly to this phenotype, since a beta2AR bearing the AT1AR tail acquired the capacity to stimulate beta-arrestin redistribution to endosomes, whereas this property was lost for an AT1AR bearing the beta2AR tail. Using beta2AR/AT1AR chimeras, we tested whether the beta2AR and AT1AR carboxyl-terminal tails, in part via their association with beta-arrestins, might regulate differences in the intracellular trafficking and resensitization patterns of these receptors. In the present study, we find that beta-arrestin formed a stable complex with the AT1AR tail in endocytic vesicles and that the internalization of this complex was dynamin dependent. Internalization of the beta2AR chimera bearing the AT1AR tail was observed in the absence of agonist and was inhibited by a dominant-negative beta-arrestin1 mutant. Agonist-independent AT1AR internalization was also observed after beta-arrestin2 overexpression. After internalization, the beta2AR, but not the AT1AR, was dephosphorylated and recycled back to the cell surface. However, the AT1AR tail prevented beta2AR dephosphorylation and recycling. In contrast, although the beta2AR-tail promoted AT1AR recycling, the chimeric receptor remained both phosphorylated and desensitized, suggesting that receptor dephosphorylation is not a property common to all receptors. In summary, we show that the carboxyl-terminal tails of GPCRs not only contribute to regulating the patterns of receptor desensitization, but also modulate receptor intracellular trafficking and resensitization patterns.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic beta-2 Receptor Agonists
  • Arrestins / genetics
  • Arrestins / metabolism*
  • Cell Line
  • Clathrin / physiology
  • Down-Regulation
  • Dynamins
  • Endocytosis*
  • GTP Phosphohydrolases / metabolism
  • Heterotrimeric GTP-Binding Proteins / metabolism
  • Humans
  • Macromolecular Substances
  • Models, Biological
  • Mutation
  • Phosphorylation
  • Precipitin Tests
  • Protein Transport
  • Receptor, Angiotensin, Type 1
  • Receptors, Adrenergic, beta-2 / genetics
  • Receptors, Adrenergic, beta-2 / metabolism*
  • Receptors, Angiotensin / agonists
  • Receptors, Angiotensin / genetics
  • Receptors, Angiotensin / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Up-Regulation
  • beta-Arrestins

Substances

  • Adrenergic beta-2 Receptor Agonists
  • Arrestins
  • Clathrin
  • Macromolecular Substances
  • Receptor, Angiotensin, Type 1
  • Receptors, Adrenergic, beta-2
  • Receptors, Angiotensin
  • Recombinant Fusion Proteins
  • beta-Arrestins
  • GTP Phosphohydrolases
  • Heterotrimeric GTP-Binding Proteins
  • Dynamins