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.

• 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.