可药性
生物信息学
G蛋白偶联受体
计算生物学
小分子
蛋白质-蛋白质相互作用
对接(动物)
药物发现
虚拟筛选
生物
受体
化学
生物信息学
生物化学
医学
基因
护理部
作者
Vincent DiGiacomo,Lien T. Nguyen,Juan B. Blanco‐Canosa,Mikel Garcia‐Marcos
标识
DOI:10.1096/fasebj.29.1_supplement.618.2
摘要
Trimeric G proteins are classically activated by G protein‐coupled receptors (GPCRs). Dysregulation of this mechanism contributes to myriad pathologies from cancer to psychiatric disorders. Indeed, GPCRs are the target for >30% of marketed drugs. G proteins can also be activated by non‐receptor proteins, however, representing a novel signaling mechanism for which no pharmacological modulators exist. Among these newly identified regulators, the GIV protein is a particularly attractive pharmacological target owing to its tight association with cancer metastasis. Moreover, GIV‐Gi coupling underlies the molecular basis of metastasis. The available structural and biochemical information indicates the interface is a tractable target for small molecules. Here, we report the development of assays compatible with high throughput screening (HTS) to monitor the direct interaction between GIV and the Gαi3 G protein. We mapped the minimal Gαi3‐binding sequence in GIV and used it to develop a fluorescence polarization‐based assay that performed robustly in a 384‐well format (Z’ ~0.7). We also developed an orthogonal counterscreen (AlphaScreen) compatible with HTS (384‐well, Z’ ~0.8). A pilot screen of the LOPAC1280 library identified 2 validated hits. Based on literature searches and in silico docking, we provide further evidence that these 2 molecules can directly target the GIV‐Gαi interface. Taken together, these results indicate that the GIV‐Gαi interface is a “druggable” protein‐protein interaction and set the framework for identifying potent anti‐metastatic agents. Support: NIH R01GM112631
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