TY - JOUR
T1 - Structure of the neurotensin receptor 1 in complex with β-arrestin 1
AU - Huang, Weijiao
AU - Masureel, Matthieu
AU - Qu, Qianhui
AU - Janetzko, John
AU - Inoue, Asuka
AU - Kato, Hideaki E.
AU - Robertson, Michael J.
AU - Nguyen, Khanh C.
AU - Glenn, Jeffrey S.
AU - Skiniotis, Georgios
AU - Kobilka, Brian K.
N1 - Funding Information:
Acknowledgements We thank M. Krawitzky and C. Adams for instrument and software access; R. Leib, F. Liu and M. Bern for discussions pertaining to mass spectrometry data analysis; G. Lam and B. Fitch for assistance with lipidomics analysis; K. Sato, Y. Sugamura and A. Inoue for plasmid construction and cell-based GPCR assays; and D. Mayer for suggestions on arrestin purification. This work was supported in part by National Institutes of Health grants R01NS028471 (B.K.K.), 1U19AI109662 (J.S.G.) and P30 CA124435 (for using the Stanford Cancer Institute Proteomics/Mass Spectrometry Shared Resource). Additional support to G.S. and B.K.K. was provided by the Mathers Foundation. B.K.K. is a Chan-Zuckerberg Biohub investigator. M.M. was supported by an American Heart Association postdoctoral fellowship (17POST33410958). J.J. is a Damon Runyon Fellow supported by the Damon Runyon Cancer Research Foundation (DRG-2318-18). A.I. was funded by the PRIME 18gm5910013 and the LEAP 18gm0010004 from the Japan Agency for Medical Research and Development (AMED) and KAKENHI 17K08264 from the Japan Society for the Promotion of Science (JSPS). H.E.K. was funded by KAKENHI 19H03163 from JSPS, The Naito Foundation, The Kurata Grants from The Hitachi Global Foundation, and Grant-in-Aid from the Tokyo Biochemical Research Foundation.
Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature Limited.
PY - 2020/3/12
Y1 - 2020/3/12
N2 - Arrestin proteins bind to active, phosphorylated G-protein-coupled receptors (GPCRs), thereby preventing G-protein coupling, triggering receptor internalization and affecting various downstream signalling pathways1,2. Although there is a wealth of structural information detailing the interactions between GPCRs and G proteins, less is known about how arrestins engage GPCRs. Here we report a cryo-electron microscopy structure of full-length human neurotensin receptor 1 (NTSR1) in complex with truncated human β-arrestin 1 (βarr1(ΔCT)). We find that phosphorylation of NTSR1 is critical for the formation of a stable complex with βarr1(ΔCT), and identify phosphorylated sites in both the third intracellular loop and the C terminus that may promote this interaction. In addition, we observe a phosphatidylinositol-4,5-bisphosphate molecule forming a bridge between the membrane side of NTSR1 transmembrane segments 1 and 4 and the C-lobe of arrestin. Compared with a structure of a rhodopsin–arrestin-1 complex, in our structure arrestin is rotated by approximately 85° relative to the receptor. These findings highlight both conserved aspects and plasticity among arrestin–receptor interactions.
AB - Arrestin proteins bind to active, phosphorylated G-protein-coupled receptors (GPCRs), thereby preventing G-protein coupling, triggering receptor internalization and affecting various downstream signalling pathways1,2. Although there is a wealth of structural information detailing the interactions between GPCRs and G proteins, less is known about how arrestins engage GPCRs. Here we report a cryo-electron microscopy structure of full-length human neurotensin receptor 1 (NTSR1) in complex with truncated human β-arrestin 1 (βarr1(ΔCT)). We find that phosphorylation of NTSR1 is critical for the formation of a stable complex with βarr1(ΔCT), and identify phosphorylated sites in both the third intracellular loop and the C terminus that may promote this interaction. In addition, we observe a phosphatidylinositol-4,5-bisphosphate molecule forming a bridge between the membrane side of NTSR1 transmembrane segments 1 and 4 and the C-lobe of arrestin. Compared with a structure of a rhodopsin–arrestin-1 complex, in our structure arrestin is rotated by approximately 85° relative to the receptor. These findings highlight both conserved aspects and plasticity among arrestin–receptor interactions.
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U2 - 10.1038/s41586-020-1953-1
DO - 10.1038/s41586-020-1953-1
M3 - Article
C2 - 31945771
AN - SCOPUS:85080034672
VL - 579
SP - 303
EP - 308
JO - Nature
JF - Nature
SN - 0028-0836
IS - 7798
ER -