title: Electrostatically-driven fast association and perdeuteration allow detection of transferred cross-relaxation for G protein-coupled receptor ligands with equilibrium dissociation constants in the high-to-low nanomolar range.
abstract: The mechanism of signal transduction mediated by G protein-coupled receptors is a subject of intense research in pharmacological and structural biology. Ligand association to the receptor constitutes a critical event in the activation process. Solution-state NMR can be amenable to high-resolution structure determination of agonist molecules in their receptor-bound state by detecting dipolar interactions in a transferred mode, even with equilibrium dissociation constants below the micromolar range. This is possible in the case of an inherent ultra-fast diffusive association of charged ligands onto a highly charged extracellular surface, and by slowing down the (1)H-(1)H cross-relaxation by perdeuterating the receptor. Here, we demonstrate this for two fatty acid molecules in interaction with the leukotriene BLT2 receptor, for which both ligands display a submicromolar affinity.
authors.affiliation: Laboratoire de Biologie Physico-Chimique des Protéines Membranaires, Institut de Biologie Physico-Chimique (FRC 550), Paris, France. laurent.catoire@ibpc.fr
authors.forename: Laurent J
authors.initials: LJ
authors.lastname: Catoire
authors.forename: Marjorie
authors.initials: M
authors.lastname: Damian
authors.forename: Marc
authors.initials: M
authors.lastname: Baaden
authors.forename: Eric
authors.initials: E
authors.lastname: Guittet
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Banères
mesh_terms: D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D015394:Molecular Structure; D043562:Receptors, G-Protein-Coupled / Q000819:agonists* / Q000737:chemistry*; D018102:Receptors, Leukotriene B4 / Q000819:agonists / Q000737:chemistry; D015398:Signal Transduction; D055672:Static Electricity
chemical_list: C485878:LTB4R2 protein, human; D043562:Receptors, G-Protein-Coupled; D018102:Receptors, Leukotriene B4