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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Heterotrimeric G proteins precouple with G protein-coupled receptors in living cells.
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Heterotrimeric G proteins precouple with G protein-coupled receptors in living cells.

机译：异源三聚体G蛋白在活细胞中与G蛋白偶联受体预偶联。

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摘要

Using fluorescence resonance energy transfer (FRET) microscopy, we investigate how heterotrimeric G proteins interact with G protein-coupled receptors (GPCRs). In the absence of receptor activation, the alpha2A adrenergic and muscarinic M4 receptors are present on the cell membrane as dimers. Furthermore, there is an interaction between the G protein subunits alpha o, beta1, and gamma2 and a number of GPCRs including M4, alpha2A, the adenosine A1 receptor, and the dopamine D2 receptor under resting conditions. The interaction between GPCRs and Galpha proteins shows specificity: there is interaction between the alpha2A receptor and Go, but little interaction between the alpha2A receptor and Gs. In contrast, the predominantly Gs-coupled prostacyclin receptor interacted with Gs, but there was little interaction between the prostacyclin receptor and Go. Inverse agonists did not change the FRET ratio, whereas the addition of agonist resulted in a modest fall. Our work suggests that GPCR dimers and the G protein heterotrimer are present at the cell membrane in the resting state in a pentameric complex.
机译：使用荧光共振能量转移（FRET）显微镜，我们调查异三聚体G蛋白如何与G蛋白偶联受体（GPCR）相互作用。在没有受体激活的情况下，α2A肾上腺素和毒蕈碱M4受体以二聚体形式存在于细胞膜上。此外，在静止条件下，G蛋白亚基alpha o，beta1和gamma2与许多GPCR（包括M4，alpha2A，腺苷A1受体和多巴胺D2受体）之间存在相互作用。 GPCR和Galpha蛋白之间的相互作用显示出特异性：alpha2A受体和Go之间存在相互作用，而alpha2A受体与Gs之间几乎没有相互作用。相反，主要是Gs偶联的前列环素受体与Gs相互作用，但是前列环素受体与Go之间几乎没有相互作用。反向激动剂没有改变FRET比率，而添加激动剂导致适度的下降。我们的工作表明，GPCR二聚体和G蛋白异源三聚体在五聚体复合物中处于静止状态时存在于细胞膜上。

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