Discovery of Small Molecules That Target the Phosphatidylinositol (3,4,5) Trisphosphate (PIP3)-Dependent Rac Exchanger 1 (P-Rex1) PIP3-Binding Site and Inhibit P-Rex1-Dependent Functions in Neutrophils
Discovery of Small Molecules That Target the Phosphatidylinositol (3,4,5) Trisphosphate (PIP3)-Dependent Rac Exchanger 1 (P-Rex1) PIP3-Binding Site and Inhibit P-Rex1-Dependent Functions in Neutrophils
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DOI：
10.1124/mol.119.117556
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发表时间：
2020-03-01
影响因子：
3.6
通讯作者：
Tesmer, John J. G.
中科院分区：
文献类型：
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作者：
Cash, Jennifer N.;Chandan, Naincy R.;Tesmer, John J. G.
磷脂酰肌醇（3，4，5）三磷酸（PIP 3）依赖性Rac交换器1（P-Rex 1）是一种Rho鸟嘌呤核苷酸交换因子，最初发现于中性粒细胞中，并受G蛋白β γ亚基和脂质PIP 3调节以响应化学引诱物。P-Rex 1也越来越多地被认为是促进乳腺癌、前列腺癌和黑色素瘤转移的作用。最近的结构、生物化学和生物学工作表明，PIP 3与P-Rex 1的普列克底物蛋白同源（PH）结构域的结合是其在细胞中活化所必需的。在这里，差示扫描荧光法用于中等通量筛选，以确定与P-Rex 1 PH结构域相互作用并阻断PIP 3结合和激活的六种小分子。其中三种化合物抑制N-甲酰基甲硫氨酰亮氨酰-苯丙氨酸诱导的人类中性粒细胞扩散以及GTPase Rac 2的激活，这两者都是P-Rex 1活性的下游效应。此外，这些化合物之一在斑马鱼模型中降低中性粒细胞速度并抑制中性粒细胞募集以响应炎症。这些结果表明，P-Rex 1的PH结构域是易处理的药物靶点，并且这些化合物可能在其它实验环境中用于抑制P-Rex 1。5）三磷酸依赖性Rac交换蛋白1（P-Rex 1）pleck-strin同源结构域具有与中性粒细胞中P-Rex 1抑制一致的作用。
Phosphatidylinositol (3,4,5) trisphosphate (PIP3)-dependent Rac exchanger 1 (P-Rex1) is a Rho guanine-nucleotide exchange factor that was originally discovered in neutrophils and is regulated by G protein beta gamma subunits and the lipid PIP3 in response to chemoattractants. P-Rex1 has also become increasingly recognized for its role in promoting metastasis of breast cancer, prostate cancer, and melanoma. Recent structural, biochemical, and biologic work has shown that binding of PIP3 to the pleckstrin homology (PH) domain of P-Rex1 is required for its activation in cells. Here, differential scanning fluorimetry was used in a medium-throughput screen to identify six small molecules that interact with the P-Rex1 PH domain and block binding of and activation by PIP3. Three of these compounds inhibit N-formylmethionylleucyl-phenylalanine induced spreading of human neutrophils as well as activation of the GTPase Rac2, both of which are downstream effects of P-Rex1 activity. Furthermore, one of these compounds reduces neutrophil velocity and inhibits neutrophil recruitment in response to inflammation in a zebrafish model. These results suggest that the PH domain of P-Rex1 is a tractable drug target and that these compounds might be useful for inhibiting P-Rex1 in other experimental contexts.SIGNIFICANCE STATEMENTA set of small molecules identified in a thermal shift screen directed against the phosphatidylinositol (3,4,5) trisphosphate-dependent Rac exchanger 1 (P-Rex1) pleck-strin homology domain has effects consistent with P-Rex1 inhibition in neutrophils.