TY  - JOUR
TI  - Expression, Purification and Crystallisation of the Adenosine A2A  Receptor Bound to an Engineered Mini G Protein
AU  - Carpenter, Byron 
AU  - Tate, Christopher Gordon
VL  - 7
IS  - 8
PY  - 2017
DA  - 2017/04/20
SP  - e2234
C1  - Bio-protocol 2017;7:e2234
DO  - 10.21769/BioProtoc.2234
UR  - https://doi.org/10.21769/BioProtoc.2234
AB  - G protein-coupled receptors (GPCRs) promote cytoplasmic signalling by activating heterotrimeric G proteins in response to extracellular stimuli such as light, hormones and nucleosides. Structure determination of GPCR–G protein complexes is central to understanding the precise mechanism of signal transduction. However, these complexes are challenging targets for structural studies due to their conformationally dynamic and inherently transient nature. We recently developed an engineered G protein, mini-Gs, which addressed these problems and allowed the formation of a stable GPCR–G protein complex. Mini-Gs facilitated the structure determination of the human adenosine A2A receptor (A2AR) in its G protein-bound conformation at 3.4 Å resolution. Here, we describe a step by step protocol for the expression and purification of A2AR, and crystallisation of the A2AR–mini-Gs complex.
KW  - Adenosine A2A receptor
KW  - A2AR
KW  - Active state
KW  - GPCR
KW  - G protein-coupled receptor
KW  - Mini G protein
KW  - Mini-Gs
KW  - G protein complex
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False