Crosslinking photosensitized by a ruthenium chelate as a tool for labeling and topographical studies of G-protein-coupled receptors

Chem Biol. 2005 Jan;12(1):15-24. doi: 10.1016/j.chembiol.2004.10.008.

Abstract

The purpose was to apply oxidative crosslinking reactions to the study of recognition and signaling mechanisms associated to G-protein-coupled receptors. Using a ruthenium chelate, Ru(bipy)(3)(2+), as photosensitizer and visible light irradiation, in the presence of ammonium persulfate, we performed fast and efficient covalent labeling of the B(2) bradykinin receptor by agonist or antagonist ligands possessing a radio-iodinated phenol moiety. The chemical and topographical specificities of these crosslinking experiments were investigated. The strategy could also be applied to the covalent labeling of the B(1) bradykinin receptor, the AT(1) angiotensin II receptor, the V(1a) vasopressin receptor and the oxytocin receptor. Interestingly, we demonstrated the possibility to covalently label the AT(1) and B(2) receptors with functionalized ligands. The potential applications of metal-chelate chemistry to receptor structural and signaling studies through intramolecular or intermolecular crosslinking are presented.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Chelating Agents / chemical synthesis
  • Chelating Agents / chemistry*
  • Ligands
  • Light
  • Molecular Sequence Data
  • Organometallic Compounds / chemical synthesis
  • Organometallic Compounds / chemistry*
  • Oxidation-Reduction
  • Peptides / chemistry
  • Photoaffinity Labels*
  • Photochemistry
  • Photosensitizing Agents / pharmacology*
  • Protein Binding / physiology
  • Receptors, G-Protein-Coupled / chemistry*
  • Receptors, G-Protein-Coupled / physiology
  • Ruthenium / chemistry*
  • Sensitivity and Specificity
  • Signal Transduction / physiology
  • Structure-Activity Relationship

Substances

  • Chelating Agents
  • Ligands
  • Organometallic Compounds
  • Peptides
  • Photoaffinity Labels
  • Photosensitizing Agents
  • Receptors, G-Protein-Coupled
  • Ruthenium