Direct binding of beta-arrestins to two distinct intracellular domains of the delta opioid receptor.
beta-Arrestins regulate opioid receptor-mediated signal transduction and play an important role in opiate-induced analgesia and tolerance/dependence. This study was carried out to measure the direct interaction between beta-arrestins and opioid receptor. Immunoprecipitation experiments demonstrated that beta-arrestin 1 physically interacts with delta opioid receptor (DOR) co-expressed in human embryonic kidney 293 cells in an agonist-enhanced manner and truncation of the carboxyl terminus of DOR partially impairs the interaction. In vitro data from glutathione-S-transferase pull-down assay showed that the carboxyl terminus (CT) and the third intracellular loop (I3L) of DOR are both capable of and either domain is sufficient for binding to beta-arrestin 1 and 2. Surface plasmon resonance determination further revealed that binding of CT and I3L of DOR to beta-arrestin is additive, suggesting these two domains bind at distinctly different sites on beta-arrestin without considerable spatial hindrance. This study demonstrated for the first time the direct binding of beta-arrestins to the two distinct domains, the carboxyl terminus and the third intracellular loop, of DOR.
Pubmed ID: 11259507 RIS Download
Amino Acid Sequence | Arrestins | Binding Sites | Cell Line | Cloning, Molecular | Cytosol | Escherichia coli | Glutathione Transferase | Humans | Kidney | Molecular Sequence Data | Peptide Fragments | Protein Conformation | Receptors, Opioid, delta | Recombinant Proteins | Transfection | beta-Arrestin 1 | beta-Arrestins