GPCR-mediated β-arrestin activation deconvoluted with single-molecule precision

Wesley B. Asher, Daniel S. Terry, G. Glenn A. Gregorio, Alem W. Kahsai, Alessandro Borgia, Bing Xie, Arnab Modak, Ying Zhu, Wonjo Jang, Alekhya Govindaraju, Li Yin Huang, Asuka Inoue, Nevin A. Lambert, Vsevolod V. Gurevich, Lei Shi, Robert J. Lefkowitz, Scott C. Blanchard, Jonathan A. Javitch

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


β-arrestins bind G protein-coupled receptors to terminate G protein signaling and to facilitate other downstream signaling pathways. Using single-molecule fluorescence resonance energy transfer imaging, we show that β-arrestin is strongly autoinhibited in its basal state. Its engagement with a phosphopeptide mimicking phosphorylated receptor tail efficiently releases the β-arrestin tail from its N domain to assume distinct conformations. Unexpectedly, we find that β-arrestin binding to phosphorylated receptor, with a phosphorylation barcode identical to the isolated phosphopeptide, is highly inefficient and that agonist-promoted receptor activation is required for β-arrestin activation, consistent with the release of a sequestered receptor C tail. These findings, together with focused cellular investigations, reveal that agonism and receptor C-tail release are specific determinants of the rate and efficiency of β-arrestin activation by phosphorylated receptor. We infer that receptor phosphorylation patterns, in combination with receptor agonism, synergistically establish the strength and specificity with which diverse, downstream β-arrestin-mediated events are directed.

Original languageEnglish (US)
Pages (from-to)1661-1675.e16
Issue number10
StatePublished - May 12 2022


  • G protein-coupled receptor
  • GPCR
  • agonist
  • arrestin
  • conformational dynamics
  • phosphorylation
  • phosphorylation barcode
  • receptor signaling
  • single-molecule FRET
  • β-arrestin

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)


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