Phospho-selective mechanisms of arrestin conformations and functions revealed by unnatural amino acid incorporation and (19)F-NMR.
Specific arrestin conformations are coupled to distinct downstream effectors, which underlie the functions of many G-protein-coupled receptors (GPCRs). Here, using unnatural amino acid incorporation and fluorine-19 nuclear magnetic resonance ((19)F-NMR) spectroscopy, we demonstrate that distinct receptor phospho-barcodes are translated to specific β-arrestin-1 conformations and direct selective signalling. With its phosphate-binding concave surface, β-arrestin-1 'reads' the message in the receptor phospho-C-tails and distinct phospho-interaction patterns are revealed by (19)F-NMR. Whereas all functional phosphopeptides interact with a common phosphate binding site and induce the movements of finger and middle loops, different phospho-interaction patterns induce distinct structural states of β-arrestin-1 that are coupled to distinct arrestin functions. Only clathrin recognizes and stabilizes GRK2-specific β-arrestin-1 conformations. The identified receptor-phospho-selective mechanism for arrestin conformation and the spacing of the multiple phosphate-binding sites in the arrestin enable arrestin to recognize plethora phosphorylation states of numerous GPCRs, contributing to the functional diversity of receptors.
Laboratory of Quantum Biophysics and Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, 100101, China.|Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.|Department of Physiology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Fan
F
Yang
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.|Department of Physiology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Xiao
X
Yu
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.
Chuan
C
Liu
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.
Chang-Xiu
CX
Qu
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.
Zheng
Z
Gong
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.
Hong-Da
HD
Liu
Laboratory of Quantum Biophysics and Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, 100101, China.
Fa-Hui
FH
Li
Department of Physiology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Hong-Mei
HM
Wang
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.|Department of Physiology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Dong-Fang
DF
He
Department of Pharmacology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Fan
F
Yi
Department of Biochemistry, University of Oxford, Oxford OX13QU, UK.
Chen
C
Song
Hefei National Laboratory for Physical Science at Microscale and School of Life Science, University of Science and Technology of China, Hefei, Anhui 230027, China.
Chang-Lin
CL
Tian
Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA.|Department of Medicine, School of Medicine, Duke University, Durham, North Carolina 27705, USA.
Kun-Hong
KH
Xiao
Laboratory of Quantum Biophysics and Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, 100101, China.
Jiang-Yun
JY
Wang
Key Laboratory Experimental Teratology of the Ministry of Education and Department of Biochemistry and Molecular Biology, Shandong University School of Medicine, 44 Wenhua Xi Road, Jinan, Shandong 250012, China.|Department of Physiology, Shandong University School of Medicine, Jinan, Shandong 250012, China.
Jin-Peng
JP
Sun
D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D015153:Blotting, Western; D002417:Cattle; D002966:Clathrin / Q000378:metabolism; D004926:Escherichia coli; D005461:Fluorine; D066235:Fluorine-19 Magnetic Resonance Imaging; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism; D054768:G-Protein-Coupled Receptor Kinases / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018613:Microscopy, Confocal; D009154:Mutation; D019906:Nuclear Magnetic Resonance, Biomolecular; D028044:Phosphate-Binding Proteins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D011487:Protein Conformation; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins
C000605253:ARRB1 protein, human; D019390:Arrestins; D002966:Clathrin; D028044:Phosphate-Binding Proteins; D010750:Phosphoproteins; D043562:Receptors, G-Protein-Coupled; D000071559:beta-Arrestin 1; D000071557:beta-Arrestins; D005461:Fluorine; C025200:3,5-difluorotyrosine; D014443:Tyrosine; C496567:GRK2 protein, human; D054769:G-Protein-Coupled Receptor Kinase 2; D054768:G-Protein-Coupled Receptor Kinases; C089176:G-protein-coupled receptor kinase 6