Agonist induced receptor internalization of neuropeptide Y receptor subtypes depends on third intracellular loop and C-terminus

Cell Signal. 2008 Oct;20(10):1740-9. doi: 10.1016/j.cellsig.2008.05.017. Epub 2008 Jun 8.

Abstract

Agonist stimulation of G-protein coupled receptors (GPCRs) results in the redistribution of the receptor from the cell surface into intracellular compartments through the process of endocytosis. Monitoring ligand-mediated internalization of GPCRs in living cells has become experimentally accessible by applying fluorescent reagents and fluorescence microscopy. By using cell lines that transiently, stably or endogenously express the human Y receptor (hYR) subtypes hY(1)R, hY(2)R, hY(4)R and hY(5)R and differently fluorescently tagged receptor proteins we were able to unravel further details concerning the internalization behavior of this multi-receptor/multi-ligand system. For the first time we could show that also the hY(2)R is internalized with a rate which is comparable to the hY(1)R and the hY(4)R. In contrast, the hY(5)R was internalized much slower and the rate remained unaffected by co-expression with other hYR subtypes. Furthermore receptor subtype co-expressing cells and selectively binding peptides revealed a receptor subtype selective internalization. By using novel hY(5)/hY(2) receptor chimera the receptor subtype dependent differences in hY receptor internalization could be identified on a molecular level.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / metabolism
  • Cell Line
  • Endocytosis*
  • Humans
  • Luminescent Proteins / metabolism
  • Molecular Sequence Data
  • Neuropeptide Y / analogs & derivatives
  • Neuropeptide Y / chemistry
  • Protein Structure, Tertiary
  • Receptors, Neuropeptide Y / agonists*
  • Receptors, Neuropeptide Y / antagonists & inhibitors
  • Receptors, Neuropeptide Y / chemistry*
  • Receptors, Neuropeptide Y / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Structure-Activity Relationship

Substances

  • Bacterial Proteins
  • Luminescent Proteins
  • Neuropeptide Y
  • Receptors, Neuropeptide Y
  • Recombinant Fusion Proteins
  • yellow fluorescent protein, Bacteria