Global insights into the fine tuning of human A2AAR conformational dynamics in a ternary complex with an engineered G protein viewed by NMR

Cell Rep. 2022 Dec 20;41(12):111844. doi: 10.1016/j.celrep.2022.111844.

Abstract

G protein-coupled receptor (GPCR) conformational plasticity enables formation of ternary signaling complexes with intracellular proteins in response to binding extracellular ligands. We investigate the dynamic process of GPCR complex formation in solution with the human A2A adenosine receptor (A2AAR) and an engineered Gs protein, mini-Gs. 2D nuclear magnetic resonance (NMR) data with uniform stable isotope-labeled A2AAR enabled a global comparison of A2AAR conformations between complexes with an agonist and mini-Gs and with an agonist alone. The two conformations are similar and show subtle differences at the receptor intracellular surface, supporting a model whereby agonist binding alone is sufficient to populate a conformation resembling the active state. However, an A2AAR "hot spot" connecting the extracellular ligand-binding pocket to the intracellular surface is observed to be highly dynamic in the ternary complex, suggesting a mechanism for allosteric connection between the bound G protein and the drug-binding pocket involving structural plasticity of the "toggle switch" tryptophan.

Keywords: A(2A) adenosine receptor; CP: Molecular biology; G(s) protein; GPCR; NMR; biophysics; mini-G(s); signaling complexes.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • GTP-Binding Proteins* / metabolism
  • Humans
  • Ligands
  • Magnetic Resonance Spectroscopy
  • Molecular Conformation
  • Protein Conformation
  • Receptor, Adenosine A2A / metabolism
  • Receptors, G-Protein-Coupled* / metabolism
  • Signal Transduction / physiology

Substances

  • GTP-Binding Proteins
  • Receptors, G-Protein-Coupled
  • Ligands
  • Receptor, Adenosine A2A