Arrestin/Clathrin Interaction LOCALIZATION OF THE CLATHRIN BINDING DOMAIN OF NONVISUAL ARRESTINS TO THE CARBOXYL TERMINUS

摘要: We have recently demonstrated that the nonvisual arrestins, β-arrestin and arrestin3, interact with high affinity stoichiometrically clathrin, we postulated this interaction mediates internalization of G protein-coupled receptors (Goodman, O. B., Jr., Krupnick, J. G., Santini, F., Gurevich, V. V., Penn, R. Gagnon, A. W., Keen, H., Benovic, L. (1996)Nature 383, 447–450). In study, localized clathrin binding domain arrestin3 using a variety approaches. Truncation mutagenesis COOH-terminal half is required for interaction. Assessment properties various glutathione S-transferase-arrestin3 fusion proteins indicated predominant contained within residues 367–385. Alanine scanning further to 371–379, site-directed critical importance both hydrophobic (Leu-373, Ile-374, Phe-376) acidic (Glu-375 Glu-377) in arrestin3/clathrin These results are complementary observation basic its arrestins L., H. (1997)J. Biol. Chem. 272, 15017–15022). Lastly, an mutant which Leu-373, Phe-376 were mutated Ala was significantly defective ability promote β2-adrenergic receptor COS-1 cells when compared wild-type arrestin3. Taken together, these implicate discrete region agonist-promoted receptor.