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Title: Mechanistic insights into allosteric regulation of the A 2A adenosine G protein-coupled receptor by physiological cations

Abstract

Cations play key roles in regulating G-protein-coupled receptors (GPCRs), although their mechanisms are poorly understood. Here, 19F NMR is used to delineate the effects of cations on functional states of the adenosine A 2A GPCR. While Na + reinforces an inactive ensemble and a partial-agonist stabilized state, Ca 2+ and Mg 2+ shift the equilibrium toward active states. Positive allosteric effects of divalent cations are more pronounced with agonist and a G-protein-derived peptide. In cell membranes, divalent cations enhance both the affinity and fraction of the high affinity agonist-bound state. Molecular dynamics simulations suggest high concentrations of divalent cations bridge specific extracellular acidic residues, bringing TM5 and TM6 together at the extracellular surface and allosterically driving open the G-protein-binding cleft as shown by rigidity-transmission allostery theory. Lastly, an understanding of cation allostery should enable the design of allosteric agents and enhance our understanding of GPCR regulation in the cellular milieu.

Authors:
 [1]; ORCiD logo [2];  [3];  [4];  [1];  [3];  [1]; ORCiD logo [5];  [2]; ORCiD logo [6]; ORCiD logo [4];  [1]
  1. Univ. of Toronto, Mississauga, ON (Canada); Univ. of Toronto, Toronto, ON (Canada)
  2. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  3. Kwansei Gakuin Univ., Nishinomiya (Japan)
  4. Univ. of California San Diego School of Medicine, La Jolla, CA (United States)
  5. Univ. of Toronto, Toronto, ON (Canada); The Hospital for Sick Children, Toronto, ON (Canada)
  6. Univ. of Toronto, Toronto, ON (Canada)
Publication Date:
Research Org.:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org.:
USDOE Laboratory Directed Research and Development (LDRD) Program
OSTI Identifier:
1435536
Report Number(s):
LA-UR-18-20653
Journal ID: ISSN 2041-1723
Grant/Contract Number:  
AC52-06NA25396
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Biological Science

Citation Formats

Ye, Libin, Neale, Chris Andrew, Sljoka, Adnan, Lyda, Brent, Pichugin, Dmitry, Tsuchimura, Nobuyuki, Larda, Sacha T., Pomes, Regis, Garcia, Angel E., Ernst, Oliver P., Sunahara, Roger K., and Prosser, R. Scott. Mechanistic insights into allosteric regulation of the A2A adenosine G protein-coupled receptor by physiological cations. United States: N. p., 2018. Web. doi:10.1038/s41467-018-03314-9.
Ye, Libin, Neale, Chris Andrew, Sljoka, Adnan, Lyda, Brent, Pichugin, Dmitry, Tsuchimura, Nobuyuki, Larda, Sacha T., Pomes, Regis, Garcia, Angel E., Ernst, Oliver P., Sunahara, Roger K., & Prosser, R. Scott. Mechanistic insights into allosteric regulation of the A2A adenosine G protein-coupled receptor by physiological cations. United States. doi:10.1038/s41467-018-03314-9.
Ye, Libin, Neale, Chris Andrew, Sljoka, Adnan, Lyda, Brent, Pichugin, Dmitry, Tsuchimura, Nobuyuki, Larda, Sacha T., Pomes, Regis, Garcia, Angel E., Ernst, Oliver P., Sunahara, Roger K., and Prosser, R. Scott. Tue . "Mechanistic insights into allosteric regulation of the A2A adenosine G protein-coupled receptor by physiological cations". United States. doi:10.1038/s41467-018-03314-9. https://www.osti.gov/servlets/purl/1435536.
@article{osti_1435536,
title = {Mechanistic insights into allosteric regulation of the A2A adenosine G protein-coupled receptor by physiological cations},
author = {Ye, Libin and Neale, Chris Andrew and Sljoka, Adnan and Lyda, Brent and Pichugin, Dmitry and Tsuchimura, Nobuyuki and Larda, Sacha T. and Pomes, Regis and Garcia, Angel E. and Ernst, Oliver P. and Sunahara, Roger K. and Prosser, R. Scott},
abstractNote = {Cations play key roles in regulating G-protein-coupled receptors (GPCRs), although their mechanisms are poorly understood. Here, 19F NMR is used to delineate the effects of cations on functional states of the adenosine A2A GPCR. While Na+ reinforces an inactive ensemble and a partial-agonist stabilized state, Ca2+ and Mg2+ shift the equilibrium toward active states. Positive allosteric effects of divalent cations are more pronounced with agonist and a G-protein-derived peptide. In cell membranes, divalent cations enhance both the affinity and fraction of the high affinity agonist-bound state. Molecular dynamics simulations suggest high concentrations of divalent cations bridge specific extracellular acidic residues, bringing TM5 and TM6 together at the extracellular surface and allosterically driving open the G-protein-binding cleft as shown by rigidity-transmission allostery theory. Lastly, an understanding of cation allostery should enable the design of allosteric agents and enhance our understanding of GPCR regulation in the cellular milieu.},
doi = {10.1038/s41467-018-03314-9},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {2018},
month = {4}
}

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    Utilization of Biased G Protein-Coupled Receptor Signaling towards Development of Safer and Personalized Therapeutics
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