TY - JOUR
T1 - β-arrestin1 and 2 exhibit distinct phosphorylation-dependent conformations when coupling to the same GPCR in living cells
AU - Haider, Raphael S.
AU - Matthees, Edda S.F.
AU - Drube, Julia
AU - Reichel, Mona
AU - Zabel, Ulrike
AU - Inoue, Asuka
AU - Chevigné, Andy
AU - Krasel, Cornelius
AU - Deupi, Xavier
AU - Hoffmann, Carsten
N1 - Acknowledgements
We thank Dr. Aurélien Rizk for his assistance in the evaluation of colocalisation in confocal image datasets. This research was supported by the European Regional Development Fund (Grant ID: EFRE HSB 2018 0019), the federal state of Thuringia and the Deutsche Forschungsgemeinschaft (GrantCRC166 ReceptorLight projectC02). J.D. is additionally funded by the University Hospital Jena IZKF (Grant ID: MSP10). A.I. was funded by the LEAP JP19gm0010004 from the Japan Agency for Medical Research and Development (AMED). A.C. was supported by the Luxembourg National Research Fund (INTER/FWO ‘Nanokine’ grant 15/10358798, INTER/FNRS grants 20/15084569, and PoC ‘Megakine’ 19/14209621), F.R.S.-FNRS-Télévie (7.8504.20).
Funding
Open Access funding enabled and organized by Projekt DEAL.
© 2022. The Author(s).
PY - 2022/9/26
Y1 - 2022/9/26
N2 - β-arrestins mediate regulatory processes for over 800 different G protein-coupled receptors (GPCRs) by adopting specific conformations that result from the geometry of the GPCR-β-arrestin complex. However, whether β-arrestin1 and 2 respond differently for binding to the same GPCR is still unknown. Employing GRK knockout cells and β-arrestins lacking the finger-loop-region, we show that the two isoforms prefer to associate with the active parathyroid hormone 1 receptor (PTH1R) in different complex configurations ("hanging" and "core"). Furthermore, the utilisation of advanced NanoLuc/FlAsH-based biosensors reveals distinct conformational signatures of β-arrestin1 and 2 when bound to active PTH1R (P-R*). Moreover, we assess β-arrestin conformational changes that are induced specifically by proximal and distal C-terminal phosphorylation and in the absence of GPCR kinases (GRKs) (R*). Here, we show differences between conformational changes that are induced by P-R* or R* receptor states and further disclose the impact of site-specific GPCR phosphorylation on arrestin-coupling and function.
AB - β-arrestins mediate regulatory processes for over 800 different G protein-coupled receptors (GPCRs) by adopting specific conformations that result from the geometry of the GPCR-β-arrestin complex. However, whether β-arrestin1 and 2 respond differently for binding to the same GPCR is still unknown. Employing GRK knockout cells and β-arrestins lacking the finger-loop-region, we show that the two isoforms prefer to associate with the active parathyroid hormone 1 receptor (PTH1R) in different complex configurations ("hanging" and "core"). Furthermore, the utilisation of advanced NanoLuc/FlAsH-based biosensors reveals distinct conformational signatures of β-arrestin1 and 2 when bound to active PTH1R (P-R*). Moreover, we assess β-arrestin conformational changes that are induced specifically by proximal and distal C-terminal phosphorylation and in the absence of GPCR kinases (GRKs) (R*). Here, we show differences between conformational changes that are induced by P-R* or R* receptor states and further disclose the impact of site-specific GPCR phosphorylation on arrestin-coupling and function.
KW - Arrestins/metabolism
KW - G-Protein-Coupled Receptor Kinases/metabolism
KW - Luciferases
KW - Parathyroid Hormone/metabolism
KW - Phosphorylation/physiology
KW - Protein Isoforms/metabolism
KW - Receptors, G-Protein-Coupled/metabolism
KW - Signal Transduction/physiology
KW - beta-Arrestin 1/genetics
KW - beta-Arrestin 2/genetics
KW - beta-Arrestins/metabolism
UR - https://www.scopus.com/pages/publications/85138653671
UR - https://pubmed.ncbi.nlm.nih.gov/36163356
U2 - 10.1038/s41467-022-33307-8
DO - 10.1038/s41467-022-33307-8
M3 - Article
C2 - 36163356
SN - 2041-1723
VL - 13
SP - 5638
JO - Nature Communications
JF - Nature Communications
IS - 1
ER -