TY - JOUR
T1 - GPCR-mediated β-arrestin activation deconvoluted with single-molecule precision
AU - Asher, Wesley B.
AU - Terry, Daniel S.
AU - Gregorio, G. Glenn A.
AU - Kahsai, Alem W.
AU - Borgia, Alessandro
AU - Xie, Bing
AU - Modak, Arnab
AU - Zhu, Ying
AU - Jang, Wonjo
AU - Govindaraju, Alekhya
AU - Huang, Li Yin
AU - Inoue, Asuka
AU - Lambert, Nevin A.
AU - Gurevich, Vsevolod V.
AU - Shi, Lei
AU - Lefkowitz, Robert J.
AU - Blanchard, Scott C.
AU - Javitch, Jonathan A.
N1 - Publisher Copyright:
© 2022 The Authors
PY - 2022/5/12
Y1 - 2022/5/12
N2 - 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.
AB - 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.
KW - G protein-coupled receptor
KW - GPCR
KW - agonist
KW - arrestin
KW - conformational dynamics
KW - phosphorylation
KW - phosphorylation barcode
KW - receptor signaling
KW - single-molecule FRET
KW - β-arrestin
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U2 - 10.1016/j.cell.2022.03.042
DO - 10.1016/j.cell.2022.03.042
M3 - Article
C2 - 35483373
AN - SCOPUS:85129971652
SN - 0092-8674
VL - 185
SP - 1661-1675.e16
JO - Cell
JF - Cell
IS - 10
ER -