Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20077
標題: Focal Adhesion Kinase與致癌基因Tpr-Met間的作用
Interaction of Focal Adhesion Kinase with Oncogene Tpr-Met
作者: 陳君梅
Chen, Chun-Mei
關鍵字: c-Met;c-Met;oncoprotein;Tpr-Met;FAK;phosphorylation;致癌蛋白;Tpr-Met;FAK;磷酸化
出版社: 生物醫學研究所
摘要: 
C-met為一種原致癌基因,其蛋白質產物為肝細胞生長因子的接受體(hepatocyte growth factor receptor, HGFR) 。Tpr-met是第一號染色體上的TPR(Translocated Promoter Region)上游區域嵌入到第七號染色體上轉譯Met kinase區域所產生的一種致癌性融合基因,它不需依賴HGF的刺激下其kinase活性持續很高,並且本身的酪氨酸呈現被磷酸化狀態。Focal adhesion kinase (FAK)是一個125 KDa的酪氨酸磷酸化激酶,存在於細胞質中,已知在integrin所調控的功能中,FAK參與了促進細胞遷移、促進細胞週期及抵抗細胞凋亡。在我的研究中,我發現Tpr-Met無論在in vivo或in vitro都可以和FAK結合並形成穩定的複合物,當Tpr-Met 上第482或489位置的酪胺酸被置換成苯丙胺酸時會破壞Tpr-Met和FAK間的結合。當我使用alkaline phosphatase處理GST-Tpr-Met時就會降低它和FAK間的結合。因此更進一步利用含Tyr-482、Tyr-489兩個酪胺酸磷酸化的phosphopeptide和Tpr-Met競爭FAK,結果指出phosphopeptide可以競爭掉Tpr-Met和FAK間的結合,證明了Tpr-Met 上第482或489位置的酪胺酸磷酸化對於其和FAK的結合的確有重要的意義。最後,我也發現無論在in vivo以及in vitro中 , FAK 的N端區域就足以和Tpr-Met直接形成穩定複合物。綜合以上結果,得到一個新的發現: Tpr-Met和FAK間的交互作用可以透過Tpr-Met上被磷酸化的Tyr-482、Tyr-489、以及FAK的N端區域來調節。

The c-met proto-oncogene encodes the hepatocyte growth factor receptor. Tpr-met is a hybrid gene resulting from a chromosomal rearrangement between chromosome 1 and chromosome 7 with its upstream region derived from the TPR locus (Translocated Promoter Region) fused to downstream sequences encoding the Met kinase. Focal adhesion kinase (FAK), a cytoplasmic protein tyrosine kinase, participates in the control of cell migration, cell cycle progression, and cell survival. In this study, I found that Tpr-Met interacted with FAK both in vivo and in vitro. Mutations at the Tyr-482 or Tyr-489 of Tpr-Met abolished this interaction. Removal of the phosphorylation of Tpr-Met by alkaline phosphatase in vitro decreased its interaction with FAK. Moreover, a synthetic phosphopeptide corresponding to the sequence surrounding the Tyr-482 and Tyr-489 was capable of blocking the interaction. Finally, the NH2-terminal domain of FAK was found to be sufficient for binding to Tpr-Met. Taken together, I found a novel interaction between Tpr-Met and FAK, which is mediated by the phosphorylated Tyr-482 and Tyr-489 of Tpr-Met and the NH2-terminal domain of FAK.
URI: http://hdl.handle.net/11455/20077
Appears in Collections:生物醫學研究所

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