[NMR paper] Determinants of ligand subtype-selectivity at ?1A-adrenoceptor revealed using Saturation Transfer Difference (STD) NMR.

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Determinants of ligand subtype-selectivity at ?1A-adrenoceptor revealed using Saturation Transfer Difference (STD) NMR.

Determinants of ligand subtype-selectivity at ?1A-adrenoceptor revealed using Saturation Transfer Difference (STD) NMR.

ACS Chem Biol. 2018 Mar 14;:

Authors: Yong KJ, Vaid TM, Shilling PJ, Wu FJ, Williams LM, Deluigi M, Plückthun A, Bathgate RA, Gooley PR, Scott DJ

Abstract
?1A- and ?1B-adrenoceptors (?1A-AR and ?1B-AR) are closely related G protein-coupled receptors (GPCRs) that modulate the cardiovascular and nervous systems in response to binding epinephrine and norepinephrine. The GPCR gene super-family is made up of numerous sub-families that, like ?1A-AR and ?1B-AR, are activated by the same endogenous agonists but may modulate different physiological processes. A major challenge in GPCR research and drug discovery is determining how compounds interact with receptors at the molecular level, especially to assist in the optimization of drug leads. Nuclear magnetic resonance spectroscopy (NMR) can provide great insight into ligand-binding epitopes, modes and kinetics. Ideally, ligand-based NMR methods require purified, well-behaved protein samples. The instability of GPCRs upon purification in detergents however, makes the application of NMR to study ligand binding challenging. Here, stabilized ?1A-AR and ?1B-AR variants were engineered using Cellular High-throughput Encapsulation, Solubilization and Screening (CHESS), allowing the analysis of ligand binding with Saturation Transfer Difference NMR (STD NMR). STD NMR was used to map the binding epitopes of epinephrine and A-61603 to both receptors, revealing the molecular determinants for the selectivity of A-61603 for ?1A-AR over ?1B-AR. The use of stabilized GPCRs for ligand-observed NMR experiments will lead to a deeper understanding of binding processes and assist structure-based drug design.


PMID: 29537256 [PubMed - as supplied by publisher]



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