Abstract
Arrestins comprise a family of signal regulators of G-protein-coupled receptors (GPCRs), which include arrestins 1 to 4. While arrestins 1 and 4 are visual arrestins dedicated to rhodopsin, arrestins 2 and 3 (Arr2 and Arr3) are β-arrestins known to regulate many nonvisual GPCRs. The dynamic and promiscuous coupling of Arr2 to nonvisual GPCRs has posed technical challenges to tackle the basis of arrestin binding to GPCRs. Here we report the structure of Arr2 in complex with neurotensin receptor 1 (NTSR1), which reveals an overall assembly that is strikingly different from the visual arrestin–rhodopsin complex by a 90° rotation of Arr2 relative to the receptor. In this new configuration, intracellular loop 3 (ICL3) and transmembrane helix 6 (TM6) of the receptor are oriented toward the N-terminal domain of the arrestin, making it possible for GPCRs that lack the C-terminal tail to couple Arr2 through their ICL3. Molecular dynamics simulation and crosslinking data further support the assembly of the Arr2‒NTSR1 complex. Sequence analysis and homology modeling suggest that the Arr2‒NTSR1 complex structure may provide an alternative template for modeling arrestin–GPCR interactions.
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Acknowledgements
The cryo-EM data were collected at Cryo-Electron Microscopy Research Center, Shanghai Institute of Material Medica. We are also grateful to the staff of the National Center for Protein Science (Shanghai) Electron Microscopy facility for instrument support. We thank the Institutional Technology Service Center of Shanghai Institute of Materia Medica, Chinese Academy of Sciences for technical assistance in mass spectrometry experiments and analysis. This work was partially supported by Ministry of Science and Technology (China) grants 2012ZX09301001, 2012CB910403, 2013CB910600, XDB08020303, and 2013ZX09507001 (to H.E.X.); Van Andel Research Institute (K.M. and H.E.X.); National Institutes of Health grants (GM127710 to H.E.X.), the 100 Talents Program of the Chinese Academy of Sciences (to X.Y.); Chinese Academy of Sciences grant (XDA12010317 to X.Y.); Natural Science Foundation of Shanghai (18ZR1447700 to X.Y.); Shanghai Sailing Program (19YF1456800 to Z.L.); China Postdoctoral Science Foundation (2019M651622 to Z.L.), National Basic Research Program of China (2017YFA0503503), NSFC (31670754), CAS (DSS-WXJZ-2018-0002, CAS-SSRC-YH-2015-01), and the CAS Major Science and Technology Infrastructure Open Research Projects to Y.C.
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W.Y. designed the expression constructs, purified the Arr2‒NTSR1 complex, prepared the final samples for negative stain and data collection toward the structures, design functional assays, performed disulfide crosslinking, and participated in figure and manuscript preparation. Z.L. performed cryo-EM data collection, processing, map refinement and figure preparation; M.J. and Y.C. performed cryo-EM sample screening, initial reconstruction and figure preparation; Y.-L.Y. and Y.Y. performed cell-based functional experiments; P.W.d.W. performed MD simulations and figure preparation; K.P., X.G., Y.H., and X.W. performed construct design and disulfide crosslinking; Y.Z. participated in EM map interpretation and figure preparation; J.G. and H.Z. performed mass spectrometry phosphorylation identification; K.M. and Y.J. participated in experimental design and manuscript editing; X.E.Z. built and refined the structure models, prepared figures and wrote the manuscript; X.Y. designed cryo-EM data collection and processing strategy, performed data processing and map refinement, and participated in manuscript editing; H.E.X. conceived and supervised the project, analyzed the structures, and wrote the manuscript.
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Yin, W., Li, Z., Jin, M. et al. A complex structure of arrestin-2 bound to a G protein-coupled receptor. Cell Res 29, 971–983 (2019). https://doi.org/10.1038/s41422-019-0256-2
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DOI: https://doi.org/10.1038/s41422-019-0256-2
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