Exploring GPCR‐arrestin interfaces with genetically encoded crosslinkers
| Autor: | Christian Ihling, Stefan Ernicke, Robert Serfling, Vsevolod V. Gurevich, Andrea Sinz, Thore Böttke, Edyta Burda, Irene Coin |
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| Rok vydání: | 2020 |
| Předmět: |
live cells
genetic structures Arrestins Cell G protein‐coupled receptor genetically encoded crosslinkers Methods & Resources Biochemistry Article Receptors G-Protein-Coupled 03 medical and health sciences 0302 clinical medicine Structural Biology DOCK Genetics medicine Arrestin Receptor Molecular Biology beta-Arrestins 030304 developmental biology G protein-coupled receptor Vasopressin receptor chemistry.chemical_classification 0303 health sciences Parathyroid Hormone Receptor 1 Articles β‐arrestins eye diseases Amino acid Cell biology medicine.anatomical_structure chemistry GPCR–arrestin complexes sense organs hormones hormone substitutes and hormone antagonists 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | EMBO Reports |
| ISSN: | 1469-3178 1469-221X |
| DOI: | 10.15252/embr.202050437 |
| Popis: | β‐arrestins (βarr1 and βarr2) are ubiquitous regulators of G protein‐coupled receptor (GPCR) signaling. Available data suggest that β‐arrestins dock to different receptors in different ways. However, the structural characterization of GPCR‐arrestin complexes is challenging and alternative approaches to study GPCR‐arrestin complexes are needed. Here, starting from the finger loop as a major site for the interaction of arrestins with GPCRs, we genetically incorporate non‐canonical amino acids for photo‐ and chemical crosslinking into βarr1 and βarr2 and explore binding topologies to GPCRs forming either stable or transient complexes with arrestins: the vasopressin receptor 2 (rhodopsin‐like), the corticotropin‐releasing factor receptor 1, and the parathyroid hormone receptor 1 (both secretin‐like). We show that each receptor leaves a unique footprint on arrestins, whereas the two β‐arrestins yield quite similar crosslinking patterns. Furthermore, we show that the method allows defining the orientation of arrestin with respect to the GPCR. Finally, we provide direct evidence for the formation of arrestin oligomers in the cell. Chemical and photo‐crosslinkers genetically incorporated into β‐arrestins allow the mapping of footprints of different receptors on the arrestin surface in living cells, and to identify intermolecular pairs of proximal amino acids in GPCR‐arrestin complexes. |
| Databáze: | OpenAIRE |
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