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

Nat Commun. 2019 Mar 19;10(1):1261. doi: 10.1038/s41467-019-09204-y.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs / physiology
  • Animals
  • Arrestin / chemistry
  • Arrestin / genetics
  • Arrestin / isolation & purification
  • Arrestin / metabolism*
  • Biological Assay
  • Cattle
  • Cell Membrane / metabolism
  • Mutation
  • Nuclear Magnetic Resonance, Biomolecular
  • Phosphorylation / physiology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Rhodopsin / chemistry
  • Rhodopsin / metabolism*
  • Rod Cell Outer Segment / metabolism
  • beta-Arrestin 1 / chemistry
  • beta-Arrestin 1 / isolation & purification
  • beta-Arrestin 1 / metabolism*
  • beta-Arrestin 2 / chemistry
  • beta-Arrestin 2 / isolation & purification
  • beta-Arrestin 2 / metabolism*

Substances

  • ARRB1 protein, human
  • ARRB2 protein, human
  • Arrestin
  • Recombinant Proteins
  • beta-Arrestin 1
  • beta-Arrestin 2
  • Rhodopsin