Crosslinking photosensitized by a ruthenium chelate as a tool for labeling and topographical studies of G-protein-coupled receptors.
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.
Duroux-Richard
Isabelle
I
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.
Vassault
Philippe
P
Subra
Guy
G
Guichou
Jean-François
JF
Richard
Eric
E
Mouillac
Bernard
B
Barberis
Claude
C
Marie
Jacky
J
Bonnafous
Jean-Claude
JC
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
D002614:Chelating Agents; D008024:Ligands; D009942:Organometallic Compounds; D010455:Peptides; D019920:Photoaffinity Labels; D017319:Photosensitizing Agents; D043562:Receptors, G-Protein-Coupled; D012428:Ruthenium