title: Analysis of temporal patterns of GPCR-β-arrestin interactions using split luciferase-fragment complementation.
abstract: We developed bioluminescence probes to detect quantitative interaction of GPCRs with arrestin isoforms β-arrestin1 and β-arrestin2 based on split luciferase complementation. Time-dependent GPCR-β-arrestin interactions showed two-types of remarkable variations that were consistent with a classification of GPCR classes. Positive charge residues in serine clusters located at the C-terminal region of GPCRs were necessary for binding to β-arrestin. This quantitative method enables elucidation of the mechanisms of different classes of GPCRs that regulate β-arrestin isoforms.
authors.affiliation: Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Tokyo 113-0033, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Hattori
authors.forename: Miho
authors.initials: M
authors.lastname: Tanaka
authors.forename: Hideo
authors.initials: H
authors.lastname: Takakura
authors.forename: Kiyono
authors.initials: K
authors.lastname: Aoki
authors.forename: Kenji
authors.initials: K
authors.lastname: Miura
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Anzai
authors.forename: Takeaki
authors.initials: T
authors.lastname: Ozawa
mesh_terms: D000595:Amino Acid Sequence; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D000071557:beta-Arrestins
chemical_list: C553443:ADRB2 protein, human; D019390:Arrestins; D018343:Receptors, Adrenergic, beta-2; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins; D008156:Luciferases