Allosteric Coupling of Drug Binding and Intracellular Signaling in the A2A Adenosine Receptor

• Published in: Cell Vol. 172; no. 1-2; pp. 68 - 80.e12
• Main Authors: Eddy, Matthew T., Lee, Ming-Yue, Gao, Zhan-Guo, White, Kate L., Didenko, Tatiana, Horst, Reto, Audet, Martin, Stanczak, Pawel, McClary, Kyle M., Han, Gye Won, Jacobson, Kenneth A., Stevens, Raymond C., Wüthrich, Kurt
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 11.01.2018; Elsevier
• Subjects: allosteric modulation; binding sites; cell membranes; drugs; G protein-coupled receptor; G-protein coupled receptors; GPCR; humans; membrane protein; NMR; nuclear magnetic resonance; nuclear magnetic resonance spectroscopy; signaling; tryptophan; X-ray diffraction; G protein-coupled receptor; NMR; signaling; GPCR; membrane protein; nuclear magnetic resonance; allosteric modulation
• ISSN: 0092-8674; 1097-4172; 1097-4172
• DOI: 10.1016/j.cell.2017.12.004

Signaling across cellular membranes, the 826 human G protein-coupled receptors (GPCRs) govern a wide range of vital physiological processes, making GPCRs prominent drug targets. X-ray crystallography provided GPCR molecular architectures, which also revealed the need for additional structural dynamics data to support drug development. Here, nuclear magnetic resonance (NMR) spectroscopy with the wild-type-like A2A adenosine receptor (A2AAR) in solution provides a comprehensive characterization of signaling-related structural dynamics. All six tryptophan indole and eight glycine backbone 15N–1H NMR signals in A2AAR were individually assigned. These NMR probes provided insight into the role of Asp522.50 as an allosteric link between the orthosteric drug binding site and the intracellular signaling surface, revealing strong interactions with the toggle switch Trp 2466.48, and delineated the structural response to variable efficacy of bound drugs across A2AAR. The present data support GPCR signaling based on dynamic interactions between two semi-independent subdomains connected by an allosteric switch at Asp522.50. [Display omitted] •Comprehensive view of GPCR signaling pathways using NMR probes•Allosteric signaling monitored by NMR probes distributed throughout A2AAR•Function-related conformational polymorphisms at intracellular A2AAR surface•Strong coupling between allosteric switch and signaling-activation motif Monitoring dynamics of GPCR signaling using stable isotope NMR reveals the path of communication enabling an allosteric response to ligand binding.