Analysis of temporal patterns of GPCR-β-arrestin interactions using split luciferase-fragment complementation

Mitsuru Hattori; Miho Tanaka; Hideo Takakura; Kiyono Aoki; Kenji Miura; Tomohiro Anzai; Takeaki Ozawa

doi:10.1039/c2mb25443c

Analysis of temporal patterns of GPCR-β-arrestin interactions using split luciferase-fragment complementation

Mol Biosyst. 2013 May;9(5):957-64. doi: 10.1039/c2mb25443c.

Authors

Mitsuru Hattori¹, Miho Tanaka, Hideo Takakura, Kiyono Aoki, Kenji Miura, Tomohiro Anzai, Takeaki Ozawa

Affiliation

¹ Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Tokyo 113-0033, Japan.

PMID: 23302795
DOI: 10.1039/c2mb25443c

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Arrestins / genetics
Arrestins / metabolism*
Blotting, Western
HEK293 Cells
Humans
Luciferases / chemistry
Luciferases / genetics
Luciferases / metabolism*
Luminescent Measurements / methods*
Molecular Sequence Data
Mutation
Protein Binding
Protein Interaction Mapping / methods*
Receptors, Adrenergic, beta-2 / genetics
Receptors, Adrenergic, beta-2 / metabolism
Receptors, G-Protein-Coupled / genetics
Receptors, G-Protein-Coupled / metabolism*
Transfection
beta-Arrestins

Substances

ADRB2 protein, human
Arrestins
Receptors, Adrenergic, beta-2
Receptors, G-Protein-Coupled
beta-Arrestins
Luciferases