Ligand modulation of sidechain dynamics in a wild-type human GPCR

Lindsay D. Clark, Igor Dikiy, Karen Chapman, Karin E.J. Rödström, James Aramini, Michael V. LeVine, George Khelashvili, Søren G.F. Rasmussen, Kevin H. Gardner, Daniel M. Rosenbaum

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

GPCRs regulate all aspects of human physiology, and biophysical studies have deepened our understanding of GPCR conformational regulation by different ligands. Yet there is no experimental evidence for how sidechain dynamics control allosteric transitions between GPCR conformations. To address this deficit, we generated samples of a wild-type GPCR (A2AR) that are deuterated apart from 1H/13C NMR probes at isoleucine δ1 methyl groups, which facilitated 1H/13C methyl TROSY NMR measurements with opposing ligands. Our data indicate that low [Na+] is required to allow large agonist-induced structural changes in A2AR, and that patterns of sidechain dynamics substantially differ between agonist (NECA) and inverse agonist (ZM241385) bound receptors, with the inverse agonist suppressing fast ps-ns timescale motions at the G protein binding site. Our approach to GPCR NMR creates a framework for exploring how different regions of a receptor respond to different ligands or signaling proteins through modulation of fast ps-ns sidechain dynamics.

Original languageEnglish (US)
Article numbere28505
JournaleLife
Volume6
DOIs
StatePublished - Oct 6 2017

Fingerprint

Nuclear magnetic resonance
Modulation
Ligands
Adenosine-5'-(N-ethylcarboxamide)
Isoleucine
Physiology
GTP-Binding Proteins
Protein Binding
Conformations
Binding Sites
Proteins
ZM 241385
Proton Magnetic Resonance Spectroscopy
Carbon-13 Magnetic Resonance Spectroscopy

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Clark, L. D., Dikiy, I., Chapman, K., Rödström, K. E. J., Aramini, J., LeVine, M. V., ... Rosenbaum, D. M. (2017). Ligand modulation of sidechain dynamics in a wild-type human GPCR. eLife, 6, [e28505]. https://doi.org/10.7554/eLife.28505

Ligand modulation of sidechain dynamics in a wild-type human GPCR. / Clark, Lindsay D.; Dikiy, Igor; Chapman, Karen; Rödström, Karin E.J.; Aramini, James; LeVine, Michael V.; Khelashvili, George; Rasmussen, Søren G.F.; Gardner, Kevin H.; Rosenbaum, Daniel M.

In: eLife, Vol. 6, e28505, 06.10.2017.

Research output: Contribution to journalArticle

Clark, LD, Dikiy, I, Chapman, K, Rödström, KEJ, Aramini, J, LeVine, MV, Khelashvili, G, Rasmussen, SGF, Gardner, KH & Rosenbaum, DM 2017, 'Ligand modulation of sidechain dynamics in a wild-type human GPCR', eLife, vol. 6, e28505. https://doi.org/10.7554/eLife.28505
Clark LD, Dikiy I, Chapman K, Rödström KEJ, Aramini J, LeVine MV et al. Ligand modulation of sidechain dynamics in a wild-type human GPCR. eLife. 2017 Oct 6;6. e28505. https://doi.org/10.7554/eLife.28505
Clark, Lindsay D. ; Dikiy, Igor ; Chapman, Karen ; Rödström, Karin E.J. ; Aramini, James ; LeVine, Michael V. ; Khelashvili, George ; Rasmussen, Søren G.F. ; Gardner, Kevin H. ; Rosenbaum, Daniel M. / Ligand modulation of sidechain dynamics in a wild-type human GPCR. In: eLife. 2017 ; Vol. 6.
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