Agonist-induced trafficking of the low-affinity formyl peptide receptor FPRL1

Cell Mol Life Sci. 2004 Jul;61(13):1684-92. doi: 10.1007/s00018-004-4116-x.

Abstract

The formyl peptide-like receptor FPRL1 is a member of the chemoattractant subfamily of G protein- coupled receptors involved in regulating leukocyte migration in inflammation. To elucidate mechanisms underlying the internalization of ligand-bound FPRL1 and possible receptor recycling, we characterized the endocytic itinerary of FPRL1. We show that agonist-triggered internalization from the plasma membrane into intracellular compartments is prevented by perturbation of clathrin-mediated endocytosis, such as expression of the dominant-negative clathrin Hub mutant, siRNA-mediated depletion of cellular clathrin and expression of a dominant-negative mutant of the large GTPase dynamin. Internalized FPRL1 co-localized with endocytosed transferrin and the small GTPases Rab4 and Rab11 in vesicular structures most resembling recycling endosomes. Recycling of FPRL1 was significantly reduced by pretreatment with PI3-kinase inhibitors. Thus, ligand-bound FPRL1 undergoes primarily clathrin-mediated and dynamin-dependent endocytosis and the receptor recycles via a rapid PI3-kinase-sensitive route as well as pathways involving perinuclear recycling endosomes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromones / pharmacology
  • Clathrin / antagonists & inhibitors
  • Clathrin / metabolism
  • Dynamins / antagonists & inhibitors
  • Dynamins / genetics
  • Dynamins / metabolism
  • Endocytosis / drug effects
  • Gene Expression Regulation, Neoplastic
  • HeLa Cells
  • Humans
  • Ligands
  • Microscopy, Fluorescence
  • Morpholines / pharmacology
  • Oligopeptides / pharmacology*
  • Phosphatidylinositol 3-Kinases / metabolism
  • Phosphoinositide-3 Kinase Inhibitors
  • Protein Transport / drug effects
  • Receptors, Formyl Peptide / agonists*
  • Receptors, Formyl Peptide / genetics
  • Receptors, Formyl Peptide / metabolism*
  • Receptors, Lipoxin / agonists*
  • Receptors, Lipoxin / genetics
  • Receptors, Lipoxin / metabolism*

Substances

  • Chromones
  • Clathrin
  • FPR2 protein, human
  • Ligands
  • Morpholines
  • Oligopeptides
  • Phosphoinositide-3 Kinase Inhibitors
  • Receptors, Formyl Peptide
  • Receptors, Lipoxin
  • Trp-Lys-Tyr-Met-Val-Met
  • 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
  • Dynamins