Illuminating G-Protein-Coupling Selectivity of GPCRs
Illuminating G-Protein-Coupling Selectivity of GPCRs
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DOI：
10.1016/j.cell.2019.04.044
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发表时间：
2019-06-13
期刊：
影响因子：
64.5
通讯作者：
Russell, Robert B.
中科院分区：
文献类型：
--
作者：
Inoue, Asuka;Raimondi, Francesco;Russell, Robert B.
异三聚体G蛋白由四个亚家族（G（s）、G（i/o）、G（q/11）和G（12/13））组成，其主要通过结合G α C末端的受体经由G蛋白偶联受体（GPCR）介导信号传导。G蛋白偶联谱支配GPCR诱导的细胞反应，但受体序列选择性决定因素仍然难以捉摸。在这里，我们系统地定量了148个GPCR和所有11个独特的G α亚基C末端之间的配体诱导的相互作用。对于每种受体，我们通过缺乏内源性G（q/11）和G（12/13）蛋白的HEK 293细胞中的转化生长因子-α（TGF-α）脱落反应探测嵌合Ga亚基活化，并通过NanoBiT-G-蛋白解离测定补充G-丙烯醛偶联谱。数据集的询问确定了跨膜结构域内部和外部的基于序列的偶联特异性特征，我们使用该特征来开发优于先前方法的偶联预测器。我们使用预测器来设计选择性地耦合到G（12）的设计者GPCR。这个不同GPCR的微调信号机制数据集是GPCR信号研究的宝贵资源。
Heterotrimetic G proteins consist of four subfamilies (G(s), G(i/o), G(q/11), and G(12/13)) that mediate signaling via G-protein-coupled receptors (GPCRs), principally by receptors binding G alpha C termini. G-protein-coupling profiles govern GPCR-induced cellular responses, yet receptor sequence selectivity determinants remain elusive. Here, we systematically quantified ligand-induced interactions between 148 GPCRs and all 11 unique G alpha subunit C termini. For each receptor, we probed chimeric Ga subunit activation via a transforming growth factor-alpha (TGF-alpha) shedding response in HEK293 cells lacking endogenous G(q/11) and G(12/13) proteins, and complemented G-prolein-coupling profiles through a NanoBiT-G-protein dissociation assay. Interrogation of the dataset identified sequence-based coupling specificity features, inside and outside the transmembrane domain, which we used to develop a coupling predictor that outperforms previous methods. We used the predictor to engineer designer GPCRs selectively coupled to G(12). This dataset of fine-tuned signaling mechanisms for diverse GPCRs is a valuable resource for research in GPCR signaling.