Membrane Mimetic-Dependence of GPCR Energy Landscapes.
Membrane Mimetic-Dependence of GPCR Energy Landscapes.
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GPCR 能量景观的膜模拟依赖性。
DOI：
10.1101/2023.10.16.562552
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Eddy,MatthewT
中科院分区：
文献类型：
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作者：
Thakur,Naveen;Ray,ArkaPrabha;Lyman,Edward;Gao,Zhan-Guo;Jacobson,KennethA;Eddy,MatthewT
我们利用可变温度的19f - nmr光谱来比较人类a2a腺苷受体（A2AAR），一种a类G蛋白偶联受体（GPCR），在19f - nmr实验中通常使用的较低温度到生理温度范围内的构象平衡。具有部分激动剂和完全激动剂的A2AAR复合物显示，随着温度的升高，完全活性构象的数量大幅增加。生理温度下的核磁共振数据更符合功能数据。这在部分激动剂的复合物中是明显的，其中活性A2AAR的数量在较低温度下几乎无法检测到，但在生理温度下变得明显。具有完全或部分激动剂的复合物的温度依赖性行为对所采用的特定膜模拟物表现出明显的敏感性。细胞信号传导实验与脂质纳米盘中A2AAR的温度依赖构象平衡相关，但与一些洗涤剂无关，强调了膜环境在研究GPCR功能中的重要性。
We leveraged variable-temperature19F-NMR spectroscopy to compare the conformational equilibria of the human A2Aadenosine receptor (A2AAR), a class A G protein-coupled receptor (GPCR), across a range of temperatures ranging from lower temperatures typically employed in19F-NMR experiments to physiological temperature. A2AAR complexes with partial agonists and full agonists showed large increases in the population of a fully active conformation with increasing temperature. NMR data measured at physiological temperature were more in line with functional data. This was pronounced for complexes with partial agonists, where the population of active A2AAR was nearly undetectable at lower temperature but became evident at physiological temperature. Temperature-dependent behavior of complexes with either full or partial agonists exhibited a pronounced sensitivity to the specific membrane mimetic employed. Cellular signaling experiments correlated with the temperature-dependent conformational equilibria of A2AAR in lipid nanodiscs but not in some detergents, underscoring the importance of the membrane environment in studies of GPCR function.