Trimethylsilyl reporter groups for NMR studies of conformational changes in G protein-coupled receptors
Trimethylsilyl reporter groups for NMR studies of conformational changes in G protein-coupled receptors
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用于 G 蛋白偶联受体构象变化 NMR 研究的三甲基甲硅烷基报告基团
DOI：
10.1002/1873-3468.13382
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发表时间：
2019-05-01
期刊：
影响因子：
3.5
通讯作者：
Liu, Dongsheng
中科院分区：
文献类型：
--
作者：
Hu, Wanhui;Wang, Huixia;Liu, Dongsheng
大分子膜蛋白，如G蛋白偶联受体(GPCRs)，由于严重的信号重叠和不利的驰豫特性，在核磁共振研究中是困难的。我们使用三甲基硅基(TMS)作为报告基团，利用接近下午0点的高强度H-1核磁共振信号，研究蛋白质的构象变化。β(2)-肾上腺素能受体被标记在位于螺旋VI和VII胞浆末端的两个半胱氨酸上。不同配体的结合导致了H-1核磁共振信号的变化，这表明螺旋VI对G蛋白特异性激活敏感，而螺旋VII对β-arrestin特异性激活敏感。因此，TMS基团是核磁共振中一个有用的报告基团，用于研究膜蛋白如GPCRs的构象变化。
Large membrane proteins such as G protein-coupled receptors (GPCRs) are difficult for NMR study due to severe signal overlaps and unfavorable relaxation properties. We used a trimethylsilyl (TMS) group as a reporter group for H-1 NMR study of conformational changes in proteins, utilizing high-intensity H-1 NMR signals near 0 p.p.m. The beta(2)-adrenergic receptor was labeled with TMS groups at two cysteines located at the cytoplasmic ends of helices VI and VII. Binding of various ligands led to changes in H-1 NMR signals, which manifested that helix VI is sensitive to G protein-specific activation, whereas helix VII is sensitive to beta-arrestin-specific activation. Thus, the TMS group is a useful reporter group in NMR for studying conformational changes in membrane proteins such as GPCRs.