G protein-coupled receptor fusion proteins in drug discovery

Curr Pharm Des. 2004;10(17):1989-2001. doi: 10.2174/1381612043384295.

Abstract

A wide range of peptides and polypeptides can be appended to either the N- or C-terminus of G protein-coupled receptors without disrupting substantially ligand binding and signal transduction. Following fusion of fluorescent proteins, reporter gene constructs or G protein alpha subunits to the C-terminal tail of a receptor high content and G protein activation assays can be employed to identify agonist ligands. Further modification of the receptor fusions to introduce enhanced levels of constitutive activity and to physically destabilise the protein allows antagonist/inverse agonists screens to be developed in parallel. Equivalent C-terminal addition of pairs of complementary, non-functional, polypeptide fragments allows the application of enzyme complementation techniques. Introduction of N-terminal tags to receptors has also allowed the introduction of novel assay techniques based on a pH-sensitive cyanine dye. These have the capacity to overcome certain limitations of GPCR-fluorescent protein fusions.

Publication types

  • Review

MeSH terms

  • Animals
  • Drug Design*
  • Green Fluorescent Proteins / genetics
  • Humans
  • Ligands
  • Protein Subunits
  • Receptors, G-Protein-Coupled / agonists*
  • Receptors, G-Protein-Coupled / antagonists & inhibitors*
  • Receptors, G-Protein-Coupled / genetics
  • Recombinant Fusion Proteins / agonists*
  • Recombinant Fusion Proteins / antagonists & inhibitors*
  • Recombinant Fusion Proteins / genetics

Substances

  • Ligands
  • Protein Subunits
  • Receptors, G-Protein-Coupled
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins