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

• Published in: Journal of biomolecular NMR Vol. 50; no. 3; pp. 191 - 195
• Main Authors: Catoire, Laurent J., Damian, Marjorie, Baaden, Marc, Guittet, Éric, Banères, Jean-Louis
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.07.2011; Springer Nature B.V; Springer Verlag
• Subjects: Biochemistry; Biochemistry, Molecular Biology; Biological and Medical Physics; Biophysics; Chemical Sciences; Communication; Humans; Life Sciences; Magnetic Resonance Spectroscopy - methods; Molecular Structure; Organic chemistry; Physics; Physics and Astronomy; Receptors, G-Protein-Coupled - agonists; Receptors, G-Protein-Coupled - chemistry; Receptors, Leukotriene B4 - agonists; Receptors, Leukotriene B4 - chemistry; Signal Transduction; Spectroscopy/Spectrometry; Static Electricity; G protein-coupled receptor; Signal transduction; Transferred NOE; Kinetics; Structural biology
• ISSN: 0925-2738; 1573-5001; 1573-5001
• DOI: 10.1007/s10858-011-9523-3

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.