The adaptor complex 2 directly interacts with the alpha 1b-adrenergic receptor and plays a role in receptor endocytosis.
Détails
ID Serval
serval:BIB_F4AC14BB2F21
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
The adaptor complex 2 directly interacts with the alpha 1b-adrenergic receptor and plays a role in receptor endocytosis.
Périodique
Journal of Biological Chemistry
ISSN
0021-9258 (Print)
ISSN-L
0021-9258
Statut éditorial
Publié
Date de publication
2003
Peer-reviewed
Oui
Volume
278
Numéro
21
Pages
19331-19340
Langue
anglais
Résumé
Using the yeast two-hybrid system, we identified the mu 2 subunit of the clathrin adaptor complex 2 as a protein interacting with the C-tail of the alpha 1b-adrenergic receptor (AR). Direct association between the alpha 1b-AR and mu 2 was demonstrated using a solid phase overlay assay. The alpha 1b-AR/mu 2 interaction occurred inside the cells, as shown by the finding that the transfected alpha 1b-AR and the endogenous mu 2 could be coimmunoprecipitated from HEK-293 cell extracts. Mutational analysis of the alpha 1b-AR revealed that the binding site for mu 2 does not involve canonical YXX Phi or dileucine motifs but a stretch of eight arginines on the receptor C-tail. The binding domain of mu 2 for the receptor C-tail involves both its N terminus and the subdomain B of its C-terminal portion. The alpha 1b-AR specifically interacted with mu 2, but not with the mu 1, mu 3, or mu 4 subunits belonging to other AP complexes. The deletion of the mu 2 binding site in the C-tail markedly decreased agonist-induced receptor internalization as demonstrated by confocal microscopy as well as by the results of a surface receptor biotinylation assay. The direct association of the adaptor complex 2 with a G protein-coupled receptor has not been reported so far and might represent a common mechanism underlying clathrin-mediated receptor endocytosis.
Mots-clé
Adaptor Protein Complex 2/chemistry, Adaptor Protein Complex 2/genetics, Adaptor Protein Complex mu Subunits/chemistry, Adaptor Protein Complex mu Subunits/genetics, Amino Acid Sequence, Animals, Binding Sites/genetics, Biotinylation, Blotting, Western, Cell Line, Clathrin/physiology, Cricetinae, Electrophoresis, Polyacrylamide Gel, Endocytosis/drug effects, Epinephrine/pharmacology, Escherichia coli/genetics, Gene Deletion, Gene Expression, Glutathione Transferase/genetics, Green Fluorescent Proteins, Humans, Immunosorbent Techniques, Luminescent Proteins/genetics, Microscopy, Confocal, Molecular Sequence Data, Mutagenesis, Polymerase Chain Reaction, Receptors, Adrenergic, alpha-1/chemistry, Receptors, Adrenergic, alpha-1/genetics, Recombinant Fusion Proteins, Transfection, Two-Hybrid System Techniques
Pubmed
Web of science
Open Access
Oui
Création de la notice
24/01/2008 12:05
Dernière modification de la notice
20/08/2019 17:21