title: Crosslinking photosensitized by a ruthenium chelate as a tool for labeling and topographical studies of G-protein-coupled receptors.
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.
authors.affiliation: Centre de Biochimie Stucturale, Centre National de la Recherche Scientifique, UMR 5048, Université Montpellier 1, Institut National de la Santé et de la Recherche Médicale, U 554, 29 rue de Navacelles, 34090 Montpellier Cedex, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Duroux-Richard
authors.forename: Philippe
authors.initials: P
authors.lastname: Vassault
authors.forename: Guy
authors.initials: G
authors.lastname: Subra
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Guichou
authors.forename: Eric
authors.initials: E
authors.lastname: Richard
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.forename: Claude
authors.initials: C
authors.lastname: Barberis
authors.forename: Jacky
authors.initials: J
authors.lastname: Marie
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Bonnafous
mesh_terms: D000595:Amino Acid Sequence; D002614:Chelating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D008024:Ligands; D008027:Light; D008969:Molecular Sequence Data; D009942:Organometallic Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry; D019920:Photoaffinity Labels; D010777:Photochemistry; D017319:Photosensitizing Agents / Q000494:pharmacology*; D011485:Protein Binding / Q000502:physiology; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000502:physiology; D012428:Ruthenium / Q000737:chemistry*; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000502:physiology; D013329:Structure-Activity Relationship
chemical_list: D002614:Chelating Agents; D008024:Ligands; D009942:Organometallic Compounds; D010455:Peptides; D019920:Photoaffinity Labels; D017319:Photosensitizing Agents; D043562:Receptors, G-Protein-Coupled; D012428:Ruthenium