Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex
Abstract The assembly of a peptide ligand, its receptor, and β-arrestin (βarr) into a ternary complex within the cell membrane is a crucial aspect of G protein-coupled receptor (GPCR) signaling. We explore this assembly by attaching fluorescent moieties to the parathyroid hormone (PTH) type 1 recept...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-12-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-54772-3 |
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| author | Jonathan Pacheco Karina A. Peña Sofya Savransky Alexandre Gidon Gerald R. V. Hammond John Janetzko Jean-Pierre Vilardaga |
| author_facet | Jonathan Pacheco Karina A. Peña Sofya Savransky Alexandre Gidon Gerald R. V. Hammond John Janetzko Jean-Pierre Vilardaga |
| author_sort | Jonathan Pacheco |
| collection | DOAJ |
| description | Abstract The assembly of a peptide ligand, its receptor, and β-arrestin (βarr) into a ternary complex within the cell membrane is a crucial aspect of G protein-coupled receptor (GPCR) signaling. We explore this assembly by attaching fluorescent moieties to the parathyroid hormone (PTH) type 1 receptor (PTH1R), using PTH as a prototypical peptide hormone, along with βarr and clathrin, and recording dual-color single-molecule imaging at the plasma membrane of live cells. Here we show that PTH1R exhibits a near-Brownian diffusion, whereas unbound hormone displays limited mobility and slow lateral diffusion at the cell surface. The formation of the PTH–PTH1R–βarr complex occurs in three sequential steps: (1) receptor and ligand collisions, (2) phosphoinositide (PIP3)-dependent recruitment and conformational change of βarr molecules at the plasma membrane, and (3) collision of most βarr molecules with the ligand-bound receptor within clathrin clusters. Our results elucidate the non-random pathway by which PTH–PTH1R–βarr complex is formed and unveil the critical role of PIP3 in regulating GPCR signaling. |
| format | Article |
| id | doaj-art-1ce330aebdeb49edb5e7abc9774e7bea |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-1ce330aebdeb49edb5e7abc9774e7bea2025-08-20T02:20:41ZengNature PortfolioNature Communications2041-17232024-12-0115111710.1038/s41467-024-54772-3Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complexJonathan Pacheco0Karina A. Peña1Sofya Savransky2Alexandre Gidon3Gerald R. V. Hammond4John Janetzko5Jean-Pierre Vilardaga6Department of Pharmacology and Chemical Biology, University of Pittsburgh School of MedicineDepartment of Pharmacology and Chemical Biology, University of Pittsburgh School of MedicineDepartment of Pharmacology and Chemical Biology, University of Pittsburgh School of MedicineDepartment of Pharmacology and Chemical Biology, University of Pittsburgh School of MedicineDepartment of Cell Biology, University of Pittsburgh School of MedicineDepartment of Molecular and Cellular Physiology, Stanford University School of MedicineDepartment of Pharmacology and Chemical Biology, University of Pittsburgh School of MedicineAbstract The assembly of a peptide ligand, its receptor, and β-arrestin (βarr) into a ternary complex within the cell membrane is a crucial aspect of G protein-coupled receptor (GPCR) signaling. We explore this assembly by attaching fluorescent moieties to the parathyroid hormone (PTH) type 1 receptor (PTH1R), using PTH as a prototypical peptide hormone, along with βarr and clathrin, and recording dual-color single-molecule imaging at the plasma membrane of live cells. Here we show that PTH1R exhibits a near-Brownian diffusion, whereas unbound hormone displays limited mobility and slow lateral diffusion at the cell surface. The formation of the PTH–PTH1R–βarr complex occurs in three sequential steps: (1) receptor and ligand collisions, (2) phosphoinositide (PIP3)-dependent recruitment and conformational change of βarr molecules at the plasma membrane, and (3) collision of most βarr molecules with the ligand-bound receptor within clathrin clusters. Our results elucidate the non-random pathway by which PTH–PTH1R–βarr complex is formed and unveil the critical role of PIP3 in regulating GPCR signaling.https://doi.org/10.1038/s41467-024-54772-3 |
| spellingShingle | Jonathan Pacheco Karina A. Peña Sofya Savransky Alexandre Gidon Gerald R. V. Hammond John Janetzko Jean-Pierre Vilardaga Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex Nature Communications |
| title | Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex |
| title_full | Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex |
| title_fullStr | Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex |
| title_full_unstemmed | Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex |
| title_short | Fast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone–GPCR–arrestin complex |
| title_sort | fast diffusing receptor collisions with slow diffusing peptide ligand assemble the ternary parathyroid hormone gpcr arrestin complex |
| url | https://doi.org/10.1038/s41467-024-54772-3 |
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