Phosphorylation-induced conformation of β2-adrenoceptor related to arrestin recruitment revealed by NMR

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Yutaro Shiraishi1, Mei Natsume1, Yutaka Kofuku1

  • 1Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo, 113-0033, Japan.

Nature Communications
|January 17, 2018
PubMed

Insights

Phosphorylation of G-protein-coupled receptors (GPCRs) by GPCR kinases causes their C-terminal regions to bind arrestin. This study reveals how phosphorylated GPCRs adopt structures that enable arrestin recognition, explaining a conserved mechanism for cellular signaling.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • G-protein-coupled receptors (GPCRs) mediate cellular responses upon agonist binding.
  • GPCR activation involves C-terminal phosphorylation by GPCR kinases, followed by arrestin binding.
  • Understanding the structural basis of GPCR-arrestin complex formation is crucial for deciphering signaling pathways.

Purpose of the Study:

  • To elucidate the structural mechanism of phosphorylated GPCR-arrestin complex formation.
  • To investigate the role of C-terminal phosphorylation in GPCR conformation and arrestin binding.
  • To identify conserved structural motifs involved in GPCR recognition by arrestin.

Main Methods:

  • Nuclear Magnetic Resonance (NMR) spectroscopy was employed.
  • Analyses were performed on phosphorylated β2-adrenoceptor (β2AR) and its complex with β-arrestin 1.
  • Experiments were conducted within lipid bilayers using nanodisc technology.

Main Results:

  • The phosphorylated C-terminal region of β2AR was observed to interact with the intracellular side of the transmembrane region or with lipids.
  • Phosphorylation induced allosteric conformational changes around key residues (M2155.54 and M2796.41) on transmembrane helices 5 and 6.
  • The phosphorylation-induced conformation closely resembles the structure associated with the β-arrestin-bound state.

Conclusions:

  • GPCR C-terminal phosphorylation induces specific conformational changes that facilitate arrestin binding.
  • A conserved structural motif in phosphorylated GPCRs allows for recognition by β-arrestin across numerous GPCR types.
  • These findings provide insights into the conserved mechanism of GPCR-arrestin interactions and downstream signaling.

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