Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein

Javier García-Nafría, Yang Lee, Xiaochen Bai, Byron Carpenter, Christopher G. Tate

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The adenosine A2A receptor (A2AR) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein GS. Here, we determine the structure by electron cryo-microscopy (cryo-EM) of A2A R at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-GS, the βγ subunits and nanobody Nb35. Most regions of the complex have a resolution of ~3.8 Å or better. Comparison with the 3.4 Å resolution crystal structure shows that the receptor and mini-GS are virtually identical and that the density of the side chains and ligand are of comparable quality. However, the cryo-EM density map also indicates regions that are flexible in comparison to the crystal structures, which unexpectedly includes regions in the ligand binding pocket. In addition, an interaction between intracellular loop 1 of the receptor and the β subunit of the G protein was observed.

Original languageEnglish (US)
Article numbere35946
JournaleLife
Volume7
DOIs
StatePublished - May 4 2018

Fingerprint

Adenosine A2A Receptors
Heterotrimeric GTP-Binding Proteins
Single-Domain Antibodies
Crystal structure
Adenosine-5'-(N-ethylcarboxamide)
Ligands
Cryoelectron Microscopy
G-Protein-Coupled Receptors
GTP-Binding Proteins
Microscopic examination
Molecules
Electrons

ASJC Scopus subject areas

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

Cite this

Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein. / García-Nafría, Javier; Lee, Yang; Bai, Xiaochen; Carpenter, Byron; Tate, Christopher G.

In: eLife, Vol. 7, e35946, 04.05.2018.

Research output: Contribution to journalArticle

García-Nafría, Javier ; Lee, Yang ; Bai, Xiaochen ; Carpenter, Byron ; Tate, Christopher G. / Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein. In: eLife. 2018 ; Vol. 7.
@article{74ae600b99034b42b8c633918430e145,
title = "Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein",
abstract = "The adenosine A2A receptor (A2AR) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein GS. Here, we determine the structure by electron cryo-microscopy (cryo-EM) of A2A R at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-GS, the βγ subunits and nanobody Nb35. Most regions of the complex have a resolution of ~3.8 {\AA} or better. Comparison with the 3.4 {\AA} resolution crystal structure shows that the receptor and mini-GS are virtually identical and that the density of the side chains and ligand are of comparable quality. However, the cryo-EM density map also indicates regions that are flexible in comparison to the crystal structures, which unexpectedly includes regions in the ligand binding pocket. In addition, an interaction between intracellular loop 1 of the receptor and the β subunit of the G protein was observed.",
author = "Javier Garc{\'i}a-Nafr{\'i}a and Yang Lee and Xiaochen Bai and Byron Carpenter and Tate, {Christopher G.}",
year = "2018",
month = "5",
day = "4",
doi = "10.7554/eLife.35946.001",
language = "English (US)",
volume = "7",
journal = "eLife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",

}

TY - JOUR

T1 - Cryo-EM structure of the adenosine A2A receptor coupled to an engineered heterotrimeric G protein

AU - García-Nafría, Javier

AU - Lee, Yang

AU - Bai, Xiaochen

AU - Carpenter, Byron

AU - Tate, Christopher G.

PY - 2018/5/4

Y1 - 2018/5/4

N2 - The adenosine A2A receptor (A2AR) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein GS. Here, we determine the structure by electron cryo-microscopy (cryo-EM) of A2A R at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-GS, the βγ subunits and nanobody Nb35. Most regions of the complex have a resolution of ~3.8 Å or better. Comparison with the 3.4 Å resolution crystal structure shows that the receptor and mini-GS are virtually identical and that the density of the side chains and ligand are of comparable quality. However, the cryo-EM density map also indicates regions that are flexible in comparison to the crystal structures, which unexpectedly includes regions in the ligand binding pocket. In addition, an interaction between intracellular loop 1 of the receptor and the β subunit of the G protein was observed.

AB - The adenosine A2A receptor (A2AR) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein GS. Here, we determine the structure by electron cryo-microscopy (cryo-EM) of A2A R at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-GS, the βγ subunits and nanobody Nb35. Most regions of the complex have a resolution of ~3.8 Å or better. Comparison with the 3.4 Å resolution crystal structure shows that the receptor and mini-GS are virtually identical and that the density of the side chains and ligand are of comparable quality. However, the cryo-EM density map also indicates regions that are flexible in comparison to the crystal structures, which unexpectedly includes regions in the ligand binding pocket. In addition, an interaction between intracellular loop 1 of the receptor and the β subunit of the G protein was observed.

UR - http://www.scopus.com/inward/record.url?scp=85049245165&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85049245165&partnerID=8YFLogxK

U2 - 10.7554/eLife.35946.001

DO - 10.7554/eLife.35946.001

M3 - Article

C2 - 29726815

AN - SCOPUS:85049245165

VL - 7

JO - eLife

JF - eLife

SN - 2050-084X

M1 - e35946

ER -