The Pyk2 FERM domain as a target to inhibit glioma migration
The Pyk2 FERM domain as a target to inhibit glioma migration
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DOI：
10.1158/1535-7163.mct-08-1055
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发表时间：
2009-06-01
影响因子：
5.7
通讯作者：
Lipinski, Christopher A.
中科院分区：
文献类型：
--
作者：
Loftus, Joseph C.;Yang, Zhongbo;Lipinski, Christopher A.
恶性胶质瘤细胞侵入周围正常脑组织，阻碍了有效的临床治疗。在这份报告中，我们研究了NH（2）末端FERM结构域在Pyk 2的促迁移功能调节中的作用。我们报告说，取代的残基，构成一个小裂缝的表面上的F3模块的FERM域不显着改变Pyk 2的表达，但导致Pyk 2磷酸化的损失。产生了特异性靶向Pyk 2 FERM结构域的单克隆抗体，命名为12 A10，并识别位于FERM结构域的F3模块的β 5C-α 1C表面上的表位。导致Pyk 2磷酸化丧失的F3模块中的氨基酸取代也抑制12 A10的结合，表明12 A10表位与在Pyk 2活性中起作用的位点重叠。12 A10与膜转运肽的缀合导致细胞内蓄积并以浓度依赖性方式抑制胶质瘤细胞迁移。12 A10的单链Fv片段在细胞内环境中表达时是稳定的，直接与Pyk 2相互作用，减少Pyk 2磷酸化，并抑制胶质瘤细胞的体外迁移。12 A10 scFv的稳定细胞内表达显著延长了神经胶质瘤异种移植模型中的存活。总之，这些数据证实了FERM结构域在调节Pyk 2活性中的核心作用，并将F3模块鉴定为抑制Pyk 2活性和抑制胶质瘤进展的新靶点。[Mol癌症治疗2009;8（6）：1505-14]
The invasion of malignant glioma cells into the surrounding normal brain precludes effective clinical treatment. In this report, we investigated the role of the NH(2)-terminal FERM domain in the regulation of the promigratory function of Pyk2. We report that the substitution of residues that constitute a small cleft on the surface of the F3 module of the FERM domain do not significantly alter Pyk2 expression but result in the loss of Pyk2 phosphorylation. A monoclonal antibody, designated 12A10, specifically targeting the Pyk2 FERM domain was generated and recognizes an epitope located on the beta 5C-alpha 1C surface of the F3 module of the FERM domain. Amino acid substitutions in the F3 module that resulted in the loss of Pyk2 phosphorylation also inhibited the binding of 12A10, suggesting that the 12A10 epitope overlaps a site that plays a role in Pyk2 activity. Conjugation of 12A10 to a membrane transport peptide led to intracellular accumulation and inhibition of glioma cell migration in a concentration-dependent manner. A single chain Fv fragment of 12A10 was stable when expressed in the intracellular environment, interacted directly with Pyk2, reduced Pyk2 phosphorylation, and inhibited glioma cell migration in vitro. Stable intracellular expression of the 12A10 scFv significantly extended survival in a glioma xenograft model. Together, these data substantiate a central role for the FERM domain in regulation of Pyk2 activity and identify the F3 module as a novel target to inhibit Pyk2 activity and inhibit glioma progression. [Mol Cancer Ther 2009;8(6):1505-14]