β-Arrestin/AP-2 Interaction in G Protein-coupled Receptor Internalization

By S. Laporte, W. Miller, Kyeong-Man Kim, M. Caron. Journal of Biological Chemistry, 2002

143 citations recorded by the source corpus.

β-Arrestins, proteins involved in the turn-off of G protein-coupled receptor (GPCR) activation, bind to the β2-adaptin subunit of the clathrin adaptor AP-2. The interaction of β2-adaptin with β-arrestin involves critical arginine residues in the C-terminal domain of β-arrestin and plays an important role in initiating clathrin-mediated endocytosis of the β2-adrenergic receptor (β2AR) (Laporte, S. A., Oakley, R. H., Holt, J. A., Barak, L. S., and Caron, M. G. (2000)J. Biol. Chem. 275, 23120–23126). However, the β-arrestin-binding site in β2-adaptin has not been identified, and little is known about the role of β-arrestin/AP-2 interaction in the endocytosis of other GPCRs. Using in vitro binding assays, we have identified two glutamate residues (Glu-849 and Glu-902) in β2-adaptin that are important in β-arrestin binding. These residues are located in the platform subdomain of the C terminus of β2-adaptin, where accessory/adapter endocytic proteins for other classes of receptors interact, distinct from the main site where clathrin interacts. The functional significance of the β-arrestin/AP-2/clathrin complex in the endocytosis of GPCRs such as the β2AR and vasopressin type II receptor was evaluated using mutant constructs of the β2-adaptin C terminus containing either the clathrin and the β-arrestin binding domains or the β-arrestin-binding domain alone. When expressed in human embryonic kidney 293 cells, both constructs acted as dominant negatives inhibiting the agonist-induced internalization of the β2AR and the vasopressin type II receptor. In addition, although the β2-adaptin construct containing both the clathrin and β-arrestin binding domains was able to block the endocytosis of transferrin receptors, a β2-adaptin construct capable of associating with β-arrestin but lacking its high affinity clathrin interaction did not interfere with transferrin receptor endocytosis. These results suggest that the interaction of β-arrestin with β2-adaptin represents a selective endocytic trigger for several members of the GPCR family.

Subfields: Cell Biology, Cellular and Molecular Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/ea9b48b72a99e5f74ad605a8b0281efd442f6cc4

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In Cellular and Molecular Neuroscience

1. J. Beaulieu
2. R. Gainetdinov

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1. C. Serhan
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Citations in the corpus

• The Physiology, Signaling, and Pharmacology of Dopamine Receptors (2011, background): “In addition to the cessation of G protein signaling, the GRK-arrestin regulatory mechanism also promotes receptor internalization from the cellular membrane through the binding of arrestins to the clathrin adaptor protein -adaptin and to clathrin itself (Laporte et al., 2002).”
• International Union of Basic and Clinical Pharmacology. LXXIII. Nomenclature for the Formyl Peptide Receptor (FPR) Family (2009, background): “It is well accepted that the -arrestins serve to link phosphorylated receptors to the components of the endocytic machinery, including clathrin and the clathrin adaptor AP2 (Goodman et al., 1996; Laporte et al., 2002).”