The Gαi-GIV binding interface is a druggable protein-protein interaction.
The Gαi-GIV binding interface is a druggable protein-protein interaction.
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DOI：
10.1038/s41598-017-08829-7
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发表时间：
2017-08-17
影响因子：
4.6
通讯作者：
Garcia-Marcos M
中科院分区：
文献类型：
--
作者：
DiGiacomo V;de Opakua AI;Papakonstantinou MP;Nguyen LT;Merino N;Blanco-Canosa JB;Blanco FJ;Garcia-Marcos M
异源三聚体G蛋白通常由GPCR的鸟嘌呤-核苷酸交换因子（GEF）活性激活。然而，一些非受体蛋白也是GEF。GIV（又名Girdin）是第一种非受体蛋白，其GEF活性归因于直接结合Gαi的明确定义的蛋白质序列。GIV表达促进转移，并且其与Gαi结合的破坏减弱癌细胞的促转移行为。尽管这表明抑制Gαi-GIV相互作用是一种有前景的治疗策略，但蛋白质-蛋白质相互作用（PPI）被认为是“可药用”靶点，需要逐个验证。在这里，我们着手研究Gαi-GIV是否是一种可药物化的PPI。我们通过计算机模拟配体筛选和化学高通量筛选（HTS）试验分别在Gαi-GIV PPI上测试了> 1，000种化合物。在二次HTS和低通量测定中确认了在两次筛选中获得的两个命中物ATA和NF 023。NF 023的结合位点，通过NMR光谱学和生化分析鉴定，与Gαi-GIV界面重叠。重要的是，NF 023不破坏Gαi-Gβγ结合，表明其对Gαi-GIV的特异性。这项工作确立了Gαi-GIV PPI作为一个可药物化的目标，并为发现这种PPI的新型抑制剂建立了概念和技术框架。
Heterotrimeric G proteins are usually activated by the guanine-nucleotide exchange factor (GEF) activity of GPCRs. However, some non-receptor proteins are also GEFs. GIV (a.k.a Girdin) was the first non-receptor protein for which the GEF activity was ascribed to a well-defined protein sequence that directly binds Gαi. GIV expression promotes metastasis and disruption of its binding to Gαi blunts the pro-metastatic behavior of cancer cells. Although this suggests that inhibition of the Gαi-GIV interaction is a promising therapeutic strategy, protein-protein interactions (PPIs) are considered poorly “druggable” targets requiring case-by-case validation. Here, we set out to investigate whether Gαi-GIV is a druggable PPI. We tested a collection of >1,000 compounds on the Gαi-GIV PPI by in silico ligand screening and separately by a chemical high-throughput screening (HTS) assay. Two hits, ATA and NF023, obtained in both screens were confirmed in secondary HTS and low-throughput assays. The binding site of NF023, identified by NMR spectroscopy and biochemical assays, overlaps with the Gαi-GIV interface. Importantly, NF023 did not disrupt Gαi-Gβγ binding, indicating its specificity toward Gαi-GIV. This work establishes the Gαi-GIV PPI as a druggable target and sets the conceptual and technical framework for the discovery of novel inhibitors of this PPI.
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影响因子：
4.8
作者：
Garcia-Marcos, Mikel;Jung, Barbara H.;Ghosh, Pradipta
通讯作者：
Ghosh, Pradipta
DOI：
10.1073/pnas.95.1.346
发表时间：
1998-01-06
影响因子：
11.1
作者：
Hohenegger, M;Waldhoer, M;Freissmuth, M
通讯作者：
Freissmuth, M
影响因子：
4.8
作者：
Garcia-Marcos, Mikel;Ghosh, Pradipta;Farquhar, Marilyn G.
通讯作者：
Farquhar, Marilyn G.
影响因子：
16.6
作者：
de Opakua AI;Parag-Sharma K;DiGiacomo V;Merino N;Leyme A;Marivin A;Villate M;Nguyen LT;de la Cruz-Morcillo MA;Blanco-Canosa JB;Ramachandran S;Baillie GS;Cerione RA;Blanco FJ;Garcia-Marcos M
通讯作者：
Garcia-Marcos M
影响因子：
4.8
作者：
Anai, M;Shojima, N;Asano, T
通讯作者：
Asano, T