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

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Abstract

The adenosine A_2A receptor (A_2AR) is a prototypical G protein-coupled receptor (GPCR) that couples to the heterotrimeric G protein G_S. Here we determine the structure by electron cryo-microscopy (cryo-EM) of A_2AR at pH 7.5 bound to the small molecule agonist NECA and coupled to an engineered heterotrimeric G protein, which contains mini-G_S, 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-G_S 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.

Data availability

Structural data have been deposited in the PDB under the accession code 6gdg and in EMDB with accession code 4390

The following data sets were generated

1. Garcia-Nafria J
2. Lee
3. Y
(2018) Cryo-EM structure of the adenosine A2A receptor bound to a miniGs heterotrimer
6GDG.

https://www.rcsb.org/

The following previously published data sets were used

1. Carpenter B
2. Nehme R
3. Warne T
4. Leslie AG
5. Tate CG
(2016) Structure of the adenosine A(2A) receptor bound to an engineered G protein.
5g53.

https://www.rcsb.org/structure/5g53

Article and author information

Author details

Javier García-Nafría

Structural Studies, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.
Yang Lee

Structural Studies, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.
Xiaochen Bai

Structural Studies, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.
Byron Carpenter

Structural Studies, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0003-1712-3528
Christopher G Tate

Structural Studies, MRC Laboratory of Molecular Biology, Cambridge, United Kingdom

For correspondence
cgt@mrc-lmb.cam.ac.uk

Competing interests
Christopher G Tate, is a consultant and shareholder of Heptares Therapeutics, and they also funded this work.

"This ORCID iD identifies the author of this article:" 0000-0002-2008-9183

Funding

Medical Research Council (U105197215)

Christopher G Tate

European Research Council (EMPSI 339995)

Christopher G Tate

Heptares Therapeutics (n/a)

Christopher G Tate

Pfizer UK (n/a)

Christopher G Tate

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.