Distinct G protein-coupled receptor phosphorylation motifs modulate arrestin affinity and activation and global conformation

Mayer, Daniel; Damberger, Fred F.; Samarasimhareddy, Mamidi; Feldmueller, Miki; Vuckovic, Ziva; Flock, Tilman; Bauer, Brian; Mutt, Eshita; Zosel, Franziska; Allain, Fr�d�ric H. T.; Standfuss, J�rg; Schertler, Gebhard F. X.; Deupi, Xavier; Sommer, Martha E.; Hurevich, Mattan; Friedler, Assaf; Veprintsev, Dmitry B.

Abstract

Cellular functions of arrestins are determined in part by the pattern of phosphorylation on the G protein-coupled receptors (GPCRs) to which arrestins bind. Despite high-resolution structural data of arrestins bound to phosphorylated receptor C-termini, the functional role of each phosphorylation site remains obscure. Here, we employ a library of synthetic phosphopeptide analogues of the GPCR rhodopsin C-terminus and determine the ability of these peptides to bind and activate arrestins using a variety of biochemical and biophysical methods. We further characterize how these peptides modulate the conformation of arrestin-1 by nuclear magnetic resonance (NMR). Our results indicate different functional classes of phosphorylation sites: ‘key sites’ required for arrestin binding and activation, an ‘inhibitory site’ that abrogates arrestin binding, and ‘modulator sites’ that influence the global conformation of arrestin. These functional motifs allow a better understanding of how different GPCR phosphorylation patterns might control how arrestin functions in the cell.

Citation

Mayer, D., Damberger, F. F., Samarasimhareddy, M., Feldmueller, M., Vuckovic, Z., Flock, T., Bauer, B., Mutt, E., Zosel, F., Allain, F. H. T., Standfuss, J., Schertler, G. F. X., Deupi, X., Sommer, M. E., Hurevich, M., Friedler, A., & Veprintsev, D. B. (2019). Distinct G protein-coupled receptor phosphorylation motifs modulate arrestin affinity and activation and global conformation. Nature Communications, 10(1), 1-14. https://doi.org/10.1038/s41467-019-09204-y

• Journal Article Type: Article
• Acceptance Date: Feb 22, 2019
• Online Publication Date: Mar 19, 2019
• Publication Date: Mar 19, 2019
• Deposit Date: Mar 16, 2020
• Publicly Available Date: Mar 16, 2020
• Journal: Nature Communications
• Electronic ISSN: 2041-1723
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 1
• Article Number: 1261
• Pages: 1-14
• DOI: https://doi.org/10.1038/s41467-019-09204-y
• Keywords: General Biochemistry, Genetics and Molecular Biology; General Physics and Astronomy; General Chemistry
• Public URL: https://nottingham-repository.worktribe.com/output/1882837
• Publisher URL: https://www.nature.com/articles/s41467-019-09204-y
• Additional Information: Received: 25 September 2018; Accepted: 22 February 2019; First Online: 19 March 2019; : The authors declare no competing interests.