期刊
ELIFE
卷 11, 期 -, 页码 -出版社
eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.73901
关键词
adenosine a2a receptor; cholesterol; G protein-coupled receptor; 19f nmr; allostery; pichia pastoris; Other
类别
资金
- CIHR Operating Grant [MOP-43998]
- Alexander Graham Bell Canada Graduate Scholarship-Doctoral from NSERC
- CREST, Japan Science and Technology Agency (JST), Japan [JPMJCR1402]
- QEII FE Beamish Graduate Scholarship in Science and Technology
Cholesterol indirectly modulates the conformational equilibria and signaling of the adenosine A(2A) receptor by influencing membrane properties.
Cholesterol is a major component of the cell membrane and commonly regulates membrane protein function. Here, we investigate how cholesterol modulates the conformational equilibria and signaling of the adenosine A(2A) receptor (A(2A)R) in reconstituted phospholipid nanodiscs. This model system conveniently excludes possible effects arising from cholesterol-induced phase separation or receptor oligomerization and focuses on the question of allostery. GTP hydrolysis assays show that cholesterol weakly enhances the basal signaling of A(2A)R while decreasing the agonist EC50. Fluorine nuclear magnetic resonance (F-19 NMR) spectroscopy shows that this enhancement arises from an increase in the receptor's active state population and a G-protein-bound precoupled state. F-19 NMR of fluorinated cholesterol analogs reveals transient interactions with A(2A)R, indicating a lack of high-affinity binding or direct allosteric modulation. The combined results suggest that the observed allosteric effects are largely indirect and originate from cholesterol-mediated changes in membrane properties, as shown by membrane fluidity measurements and high-pressure NMR.
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