The beta2-adrenergic receptor/betaarrestin complex recruits the clathrin adaptor AP-2 during endocytosis

Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3712-7. doi: 10.1073/pnas.96.7.3712.

Abstract

betaarrestins mediate the desensitization of the beta2-adrenergic receptor (beta2AR) and many other G protein-coupled receptors (GPCRs). Additionally, betaarrestins initiate the endocytosis of these receptors via clathrin coated-pits and interact directly with clathrin. Consequently, it has been proposed that betaarrestins serve as clathrin adaptors for the GPCR family by linking these receptors to clathrin lattices. AP-2, the heterotetrameric clathrin adaptor protein, has been demonstrated to mediate the internalization of many types of plasma membrane proteins other than GPCRs. AP-2 interacts with the clathrin heavy chain and cytoplasmic domains of receptors such as those for epidermal growth factor and transferrin. In the present study we demonstrate the formation of an agonist-induced multimeric complex containing a GPCR, betaarrestin 2, and the beta2-adaptin subunit of AP-2. beta2-Adaptin binds betaarrestin 2 in a yeast two-hybrid assay and coimmunoprecipitates with betaarrestins and beta2AR in an agonist-dependent manner in HEK-293 cells. Moreover, beta2-adaptin translocates from the cytosol to the plasma membrane in response to the beta2AR agonist isoproterenol and colocalizes with beta2AR in clathrin-coated pits. Finally, expression of betaarrestin 2 minigene constructs containing the beta2-adaptin interacting region inhibits beta2AR endocytosis. These findings point to a role for AP-2 in GPCR endocytosis, and they suggest that AP-2 functions as a clathrin adaptor for the endocytosis of diverse classes of membrane receptors.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Protein Complex alpha Subunits
  • Adaptor Protein Complex beta Subunits / chemistry
  • Adaptor Protein Complex beta Subunits / genetics
  • Adaptor Protein Complex beta Subunits / physiology*
  • Adaptor Proteins, Vesicular Transport
  • Amino Acid Sequence
  • Animals
  • Arrestins / genetics
  • Arrestins / physiology*
  • COS Cells
  • Cell Line
  • Cloning, Molecular
  • Endocytosis / physiology*
  • Green Fluorescent Proteins
  • Humans
  • Luminescent Proteins / biosynthesis
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Macromolecular Substances
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / physiology*
  • Molecular Sequence Data
  • Receptors, Adrenergic, beta-2 / genetics
  • Receptors, Adrenergic, beta-2 / physiology*
  • Recombinant Fusion Proteins / biosynthesis
  • Saccharomyces cerevisiae
  • Transfection
  • beta-Arrestins

Substances

  • AP2B1 protein, human
  • Adaptor Protein Complex alpha Subunits
  • Adaptor Protein Complex beta Subunits
  • Adaptor Proteins, Vesicular Transport
  • Arrestins
  • Luminescent Proteins
  • Macromolecular Substances
  • Membrane Proteins
  • Receptors, Adrenergic, beta-2
  • Recombinant Fusion Proteins
  • beta-Arrestins
  • Green Fluorescent Proteins