Structural studies of phosphorylation-dependent interactions between the V2R receptor and arrestin-2

• Published in: Nature communications Vol. 12; no. 1; pp. 2396 - 16
• Main Authors: He, Qing-Tao, Xiao, Peng, Huang, Shen-Ming, Jia, Ying-Li, Zhu, Zhong-Liang, Lin, Jing-Yu, Yang, Fan, Tao, Xiao-Na, Zhao, Ru-Jia, Gao, Feng-Yuan, Niu, Xiao-Gang, Xiao, Kun-Hong, Wang, Jiangyun, Jin, Changwen, Sun, Jin-Peng, Yu, Xiao
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 22.04.2021; Nature Publishing Group; Nature Portfolio
• Subjects: 631/45/535/1266; 631/535/1266; 82/6; 82/80; 82/83; Arrestin; beta-Arrestin 1 - genetics; beta-Arrestin 1 - isolation & purification; beta-Arrestin 1 - metabolism; beta-Arrestin 1 - ultrastructure; Crystal structure; Crystallography, X-Ray; Fluorescence resonance energy transfer; Functionals; G protein-coupled receptors; Genetic analysis; Genetic code; HEK293 Cells; Humanities and Social Sciences; Humans; multidisciplinary; Mutation; NMR; Nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular; Phosphopeptides - chemistry; Phosphopeptides - metabolism; Phosphorylation; Protein Conformation, alpha-Helical; Protein Domains - genetics; Receptors; Receptors, Vasopressin - chemistry; Receptors, Vasopressin - metabolism; Receptors, Vasopressin - ultrastructure; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Recombinant Proteins - ultrastructure; Science; Science (multidisciplinary); Spectrum analysis; Structure-function relationships; Vasopressin; Vasopressin V2 receptors
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-22731-x

Arrestins recognize different receptor phosphorylation patterns and convert this information to selective arrestin functions to expand the functional diversity of the G protein-coupled receptor (GPCR) superfamilies. However, the principles governing arrestin-phospho-receptor interactions, as well as the contribution of each single phospho-interaction to selective arrestin structural and functional states, are undefined. Here, we determined the crystal structures of arrestin2 in complex with four different phosphopeptides derived from the vasopressin receptor-2 (V2R) C-tail. A comparison of these four crystal structures with previously solved Arrestin2 structures demonstrated that a single phospho-interaction change results in measurable conformational changes at remote sites in the complex. This conformational bias introduced by specific phosphorylation patterns was further inspected by FRET and 1 H NMR spectrum analysis facilitated via genetic code expansion. Moreover, an interdependent phospho-binding mechanism of phospho-receptor-arrestin interactions between different phospho-interaction sites was unexpectedly revealed. Taken together, our results provide evidence showing that phospho-interaction changes at different arrestin sites can elicit changes in affinity and structural states at remote sites, which correlate with selective arrestin functions. The interaction between a GPCR, such as the vasopressin receptor-2 (V2R), and arrestin depends on the receptors’ phosphorylation pattern. Here authors use FRET and NMR to analyze the phosphorylation patterns of the V2R-arrestin complex and show that phospho-interactions are the key determinants of selective arrestin conformational states and correlated functions.