Skip to main content
Journal cover image

Visualization of arrestin recruitment by a G-protein-coupled receptor.

Publication ,  Journal Article
Shukla, AK; Westfield, GH; Xiao, K; Reis, RI; Huang, L-Y; Tripathi-Shukla, P; Qian, J; Li, S; Blanc, A; Oleskie, AN; Dosey, AM; Su, M ...
Published in: Nature
August 14, 2014

G-protein-coupled receptors (GPCRs) are critically regulated by β-arrestins, which not only desensitize G-protein signalling but also initiate a G-protein-independent wave of signalling. A recent surge of structural data on a number of GPCRs, including the β2 adrenergic receptor (β2AR)-G-protein complex, has provided novel insights into the structural basis of receptor activation. However, complementary information has been lacking on the recruitment of β-arrestins to activated GPCRs, primarily owing to challenges in obtaining stable receptor-β-arrestin complexes for structural studies. Here we devised a strategy for forming and purifying a functional human β2AR-β-arrestin-1 complex that allowed us to visualize its architecture by single-particle negative-stain electron microscopy and to characterize the interactions between β2AR and β-arrestin 1 using hydrogen-deuterium exchange mass spectrometry (HDX-MS) and chemical crosslinking. Electron microscopy two-dimensional averages and three-dimensional reconstructions reveal bimodal binding of β-arrestin 1 to the β2AR, involving two separate sets of interactions, one with the phosphorylated carboxy terminus of the receptor and the other with its seven-transmembrane core. Areas of reduced HDX together with identification of crosslinked residues suggest engagement of the finger loop of β-arrestin 1 with the seven-transmembrane core of the receptor. In contrast, focal areas of raised HDX levels indicate regions of increased dynamics in both the N and C domains of β-arrestin 1 when coupled to the β2AR. A molecular model of the β2AR-β-arrestin signalling complex was made by docking activated β-arrestin 1 and β2AR crystal structures into the electron microscopy map densities with constraints provided by HDX-MS and crosslinking, allowing us to obtain valuable insights into the overall architecture of a receptor-arrestin complex. The dynamic and structural information presented here provides a framework for better understanding the basis of GPCR regulation by arrestins.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

Nature

DOI

EISSN

1476-4687

Publication Date

August 14, 2014

Volume

512

Issue

7513

Start / End Page

218 / 222

Location

England

Related Subject Headings

  • beta-Arrestins
  • beta-Arrestin 1
  • Sf9 Cells
  • Receptors, G-Protein-Coupled
  • Receptors, Adrenergic, beta-2
  • Protein Structure, Quaternary
  • Models, Molecular
  • General Science & Technology
  • GTP-Binding Proteins
  • Arrestins
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Shukla, A. K., Westfield, G. H., Xiao, K., Reis, R. I., Huang, L.-Y., Tripathi-Shukla, P., … Lefkowitz, R. J. (2014). Visualization of arrestin recruitment by a G-protein-coupled receptor. Nature, 512(7513), 218–222. https://doi.org/10.1038/nature13430
Shukla, Arun K., Gerwin H. Westfield, Kunhong Xiao, Rosana I. Reis, Li-Yin Huang, Prachi Tripathi-Shukla, Jiang Qian, et al. “Visualization of arrestin recruitment by a G-protein-coupled receptor.Nature 512, no. 7513 (August 14, 2014): 218–22. https://doi.org/10.1038/nature13430.
Shukla AK, Westfield GH, Xiao K, Reis RI, Huang L-Y, Tripathi-Shukla P, et al. Visualization of arrestin recruitment by a G-protein-coupled receptor. Nature. 2014 Aug 14;512(7513):218–22.
Shukla, Arun K., et al. “Visualization of arrestin recruitment by a G-protein-coupled receptor.Nature, vol. 512, no. 7513, Aug. 2014, pp. 218–22. Pubmed, doi:10.1038/nature13430.
Shukla AK, Westfield GH, Xiao K, Reis RI, Huang L-Y, Tripathi-Shukla P, Qian J, Li S, Blanc A, Oleskie AN, Dosey AM, Su M, Liang C-R, Gu L-L, Shan J-M, Chen X, Hanna R, Choi M, Yao XJ, Klink BU, Kahsai AW, Sidhu SS, Koide S, Penczek PA, Kossiakoff AA, Woods VL, Kobilka BK, Skiniotis G, Lefkowitz RJ. Visualization of arrestin recruitment by a G-protein-coupled receptor. Nature. 2014 Aug 14;512(7513):218–222.
Journal cover image

Published In

Nature

DOI

EISSN

1476-4687

Publication Date

August 14, 2014

Volume

512

Issue

7513

Start / End Page

218 / 222

Location

England

Related Subject Headings

  • beta-Arrestins
  • beta-Arrestin 1
  • Sf9 Cells
  • Receptors, G-Protein-Coupled
  • Receptors, Adrenergic, beta-2
  • Protein Structure, Quaternary
  • Models, Molecular
  • General Science & Technology
  • GTP-Binding Proteins
  • Arrestins