Rational design and characterization of a Rac GTPase-specific small molecule inhibitor
Rational design and characterization of a Rac GTPase-specific small molecule inhibitor
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DOI：
10.1073/pnas.0307512101
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发表时间：
2004-05-18
影响因子：
11.1
通讯作者：
Zheng, Y
中科院分区：
文献类型：
--
作者：
Gao, Y;Dickerson, JB;Zheng, Y
由Rho家族GTP酶介导的信号传导途径涉及细胞生物学的许多方面。这些途径的特异性部分是通过DbI家族鸟嘌呤核苷酸交换因子（GEF）与其Rho GTdR底物之间的选择性相互作用实现的。在这里，我们报告了第一代小分子Rac GTdR抑制剂，靶向Rac激活GEF。化合物NSC 23766通过基于结构的化合物虚拟筛选来鉴定，所述化合物适合已知对GEF规范至关重要的Rac 1表面凹槽。在体外，它可以有效地抑制Rac 1的结合和激活的Rac特异性GEF Trio或Tiam 1在剂量依赖性的方式，而不干扰密切相关的Cdc 42或RhoA的结合或激活其各自的GEF或与Rac 1的相互作用与BcrGAP或效应PAK 1。在细胞中，它有效地阻断血清或血小板衍生生长因子诱导的Rac 1激活和板状伪足形成，而不影响内源性Cdc 42或RhoA的活性。此外，该化合物减少Trio或Tiam 1，但不减少Vav、Lbc、Intersectin或组成型活性Rac 1突变体刺激的细胞生长，并抑制Trio、Tiaml或Ras诱导的细胞转化。当应用于人前列腺癌PC-3细胞时，它能够抑制需要内源性Rac 1活性的增殖、锚定非依赖性生长和侵袭表型。因此，NSC 23766构成Rac特异性小分子抑制剂，其可用于研究Rac在各种细胞功能中的作用，并逆转与Rac失调相关的肿瘤细胞表型。
The signaling pathways mediated by Rho family GTPases have been implicated in many aspects of cell biology. The specificity of the pathways is achieved in part by the selective interaction between DbI family guanine nucleotide exchange factors (GEFs) and their Rho GTPase substrates. Here, we report a first-generation small-molecule inhibitor of Rac GTPase targeting Rac activation by GEF. The chemical compound NSC23766 was identified by a structure-based virtual screening of compounds that fit into a surface groove of Rac1 known to be critical for GEF specification. In vitro it could effectively inhibit Rac1 binding and activation by the Rac-specific GEF Trio or Tiam1 in a dose-dependent manner without interfering with the closely related Cdc42 or RhoA binding or activation by their respective GEFs or with Rac1 interaction with BcrGAP or effector PAK1. In cells, it potently blocked serum or platelet-derived growth factor-induced Rac1 activation and lamellipodia formation without affecting the activity of endogenous Cdc42 or RhoA. Moreover, this compound reduced Trio or Tiam1 but not Vav, Lbc, Intersectin, or a constitutively active Rac1 mutant-stimulated cell growth and suppressed Trio, Tiaml, or Ras-induced cell transformation. When applied to human prostate cancer PC-3 cells, it was able to inhibit the proliferation, anchorage-independent growth and invasion phenotypes that require the endogenous Rac1 activity. Thus, NSC23766 constitutes a Rac-specific small-molecule inhibitor that could be useful to study the role of Rac in various cellular functions and to reverse tumor cell phenotypes associated with Rac deregulation.