Allosteric Inhibitors of the Eya2 Phosphatase Are Selective and Inhibit Eya2-mediated Cell Migration
Allosteric Inhibitors of the Eya2 Phosphatase Are Selective and Inhibit Eya2-mediated Cell Migration
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DOI：
10.1074/jbc.m114.566729
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发表时间：
2014-06-06
影响因子：
4.8
通讯作者：
Zhao, Rui
中科院分区：
文献类型：
--
作者：
Krueger, Aaron B.;Drasin, David J.;Zhao, Rui
Eya蛋白是Six家族转录因子的必需共激活因子，并且含有属于磷酸酶的卤酸脱卤酶家族的独特酪氨酸磷酸酶结构域。Eya的磷酸酶活性对于Six 1靶基因的一个子集的转录是重要的，并且还指导细胞在DNA损伤时进行修复而不是凋亡途径。此外，Eya磷酸酶活性已被证明介导乳腺癌细胞的转化、侵袭、迁移和转移，使其成为乳腺癌的潜在新药靶点。我们先前已经鉴定了一类特异性抑制Eya 2磷酸酶的N-亚芳基苯甲酰肼化合物。在此，我们证明这些化合物是可逆的抑制剂，选择性地抑制Eya 2的磷酸酶活性，但不抑制Eya 3。我们的诱变结果表明，这类化合物不结合的活性位点和结合不需要与Mg 2+的配位。此外，这些化合物可能结合在活性位点的相对面上的位点内，并作为变构抑制剂起作用。我们还证明了这类化合物抑制Eya 2磷酸酶介导的细胞迁移，为这些分子被开发成化学探针以了解Eya 2磷酸酶的特定功能奠定了基础，并作为开发Eya 2磷酸酶特异性抗癌药物的原型。
Eya proteins are essential co-activators of the Six family of transcription factors and contain a unique tyrosine phosphatase domain belonging to the haloacid dehalogenase family of phosphatases. The phosphatase activity of Eya is important for the transcription of a subset of Six1-target genes, and also directs cells to the repair rather than apoptosis pathway upon DNA damage. Furthermore, Eya phosphatase activity has been shown to mediate transformation, invasion, migration, and metastasis of breast cancer cells, making it a potential new drug target for breast cancer. We have previously identified a class of N-arylidenebenzohydrazide compounds that specifically inhibit the Eya2 phosphatase. Herein, we demonstrate that these compounds are reversible inhibitors that selectively inhibit the phosphatase activity of Eya2, but not Eya3. Our mutagenesis results suggest that this class of compounds does not bind to the active site and the binding does not require the coordination with Mg2+. Moreover, these compounds likely bind within a site on the opposite face of the active site, and function as allosteric inhibitors. We also demonstrate that this class of compounds inhibits Eya2 phosphatase-mediated cell migration, setting the foundation for these molecules to be developed into chemical probes for understanding the specific function of the Eya2 phosphatase and to serve as a prototype for the development of Eya2 phosphatase specific anti-cancer drugs.