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.
N1 - Funding Information:
This work was funded by a grant from the European Research Council (EMPSI 339995) Heptares Therapeutics Ltd, Pfizer and core funding from the Medical Research Council [MRC U105197215]. We thank Rishi Matadeen and Kasim Sader for their help with cryo-EM data collection, Christos Savva for his help in using the VPP and Wim Hagen for the SerialEM single-particle data collection script. We also thank Rafael Fernandez-Leiro, Sjors Scheres and Paula da Fonseca for useful discussions.
Publisher Copyright:
© García-Nafría et al.
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.
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U2 - 10.7554/eLife.35946.001
DO - 10.7554/eLife.35946.001
M3 - Article
C2 - 29726815
AN - SCOPUS:85049245165
SN - 2050-084X
VL - 7
JO - eLife
JF - eLife
M1 - e35946
ER -