Inhibition of chemoattractant N-formyl peptide receptor trafficking by active arrestins

Traffic. 2005 Feb;6(2):87-99. doi: 10.1111/j.1600-0854.2004.00248.x.

Abstract

Recent studies have highlighted the emergence of a class of G protein-coupled receptors that are internalized in an arrestin-independent manner. In addition to demonstrating that the N-formyl peptide receptor belongs in this family, we have recently shown that recycling of the receptor requires the presence of arrestins. To further elucidate mechanisms of arrestin-dependent regulation of G protein-coupled receptor processing, we examined the effects of altering the receptor-arrestin complex on ternary complex formation and cellular trafficking of the N-formyl peptide receptor by studying two active arrestin-2 mutants (truncated arrestin-2 [1-382], and arrestin-2 I386A, V387A, F388A). Complexes between the N-formyl peptide receptor and active arrestins exhibited higher affinity in vitro than the complex between the N-formyl peptide receptor and wild-type arrestin and furthermore were observed in vivo by colocalization studies using confocal microscopy. To assess the effects of these altered interactions on receptor trafficking, we demonstrated that active, but not wild-type, arrestin expression retards N-formyl peptide receptor internalization. Furthermore, expression of arrestin-2 I386A/V387A/F388A but not arrestin-2 [1-382] inhibited recycling of the N-formyl peptide receptor, reflecting an expanded role for arrestins in G protein-coupled receptor processing and trafficking. Whereas the extent of N-formyl peptide receptor phosphorylation had no effect on the inhibition of internalization, N-formyl peptide receptor recycling was restored when the receptor was only partially phosphorylated. These results indicate not only that a functional interaction between receptor and arrestin is required for recycling of certain G protein-coupled receptors, such as the N-formyl peptide receptor, but that the pattern of receptor phosphorylation further regulates this process.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Arrestins / agonists
  • Arrestins / chemistry
  • Arrestins / genetics
  • Arrestins / metabolism*
  • Arrestins / pharmacology
  • Endocytosis
  • Fluorescent Dyes
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Kinetics
  • Ligands
  • Microscopy, Confocal
  • Phosphorylation
  • Point Mutation
  • Protein Transport
  • Quinolinium Compounds
  • Receptors, Formyl Peptide / antagonists & inhibitors*
  • Spectrometry, Fluorescence
  • U937 Cells

Substances

  • Alexa fluor 546
  • Arrestins
  • Fluorescent Dyes
  • Ligands
  • Quinolinium Compounds
  • Receptors, Formyl Peptide
  • Green Fluorescent Proteins