Sun Zhongya, Zhang Hao, Zhang Yuanyuan, Liao Liping, Zhou Wen, Zhang Fengcai, Lian Fulin, Huang Jing, Xu Pan, Zhang Rukang, Lu Wenchao, Zhu Mingrui, Tao Hongru, Yang Feng, Ding Hong, Chen Shijie, Yue Liyan, Zhou Bing, Zhang Naixia, Tan Minjia, Jiang Hualiang, Chen Kaixian, Liu Bo, Liu Chuanpeng, Dang Yongjun, Luo Cheng
School of Life Science and Technology Harbin Institute of Technology Harbin 150001 China.
Drug Discovery and Design Center State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai 201203 China.
Adv Sci (Weinh). 2020 May 13;7(14):2000098. doi: 10.1002/advs.202000098. eCollection 2020 Jul.
The Rho family GTPases are crucial drivers of tumor growth and metastasis. However, it is difficult to develop GTPases inhibitors due to a lack of well-characterized binding pockets for compounds. Here, through molecular dynamics simulation of the RhoA protein, a groove around cysteine 107 (Cys107) that is relatively well-conserved within the Rho family is discovered. Using a combined strategy, the novel inhibitor DC-Rhoin is discovered, which disrupts interaction of Rho proteins with guanine nucleotide exchange factors (GEFs) and guanine nucleotide dissociation inhibitors (GDIs). Crystallographic studies reveal that the covalent binding of DC-Rhoin to the Cys107 residue stabilizes and captures a novel allosteric pocket. Moreover, the derivative compound DC-Rhoin04 inhibits the migration and invasion of cancer cells, through targeting this allosteric pocket of RhoA. The study reveals a novel allosteric regulatory site within the Rho family, which can be exploited for anti-metastasis drug development, and also provides a novel strategy for inhibitor discovery toward "undruggable" protein targets.
Rho家族GTP酶是肿瘤生长和转移的关键驱动因素。然而，由于缺乏特征明确的化合物结合口袋，开发GTP酶抑制剂具有一定难度。在此，通过对RhoA蛋白进行分子动力学模拟，发现了一个围绕半胱氨酸107（Cys107）的凹槽，该凹槽在Rho家族中相对保守。采用联合策略，发现了新型抑制剂DC-Rhoin，它可破坏Rho蛋白与鸟嘌呤核苷酸交换因子（GEFs）和鸟嘌呤核苷酸解离抑制剂（GDIs）的相互作用。晶体学研究表明，DC-Rhoin与Cys107残基的共价结合稳定并捕获了一个新的变构口袋。此外，衍生物化合物DC-Rhoin04通过靶向RhoA的这个变构口袋，抑制癌细胞的迁移和侵袭。该研究揭示了Rho家族内一个新的变构调节位点，可用于抗转移药物开发，也为针对“不可成药”蛋白靶点的抑制剂发现提供了新策略。
