臺灣博碩士論文加值系統

EPS8 (EGFR pathway substrate no.8)是由Dr. Di Fiore實驗室於1993年所發現的一個蛋白質，具有97kDa及68kDa二種isoforms，但以p97Eps8為major form。它會受receptor tyrosine kinases (如:EGFR，PDGFR，FGFR)及non-receptor tyrosine kinase (如:Src)的磷酸化。先前我們實驗室指出在v-Src所造成的癌細胞中，Eps8蛋白質的表現量及tyrosine磷酸化的情形均有增加的現象。另外之前實驗室藉由tryptic phosphopeptide maps發現，在in vitro 下p97Eps8有三個tyrosine的位置會受到Src的磷酸化，而其中一個主要的磷酸化位置就是Tyr-45，另外二個的位置是存於p68Eps8上。因此我們利用Site-directed mutagenesis及Mass spectrophotometer來分析細胞內Src磷酸化p68Eps8的位置。我們發現到Tyr-524及Tyr-674是細胞Src磷酸化Eps8的另外二個磷酸化位置。以in vitro kinase reaction也證明Tyr-524及Tyr-674是Src磷酸化的位置。

Eps8 was initially identified as a substrate of EGF receptor tyrosine kinase. Our previous studies have indicated that Eps8 could also be phosphorylated by both v-Src and active c-Src. Interestingly, both the expression and tyrosyl phosphorylation of Eps8 are enhanced in v-Src transformed cells. Thus, protein phosphorylation and expression of Eps8 may contribute to Src-mediated mitogenesis and oncogenesis. Previously, we have observed at least three potential Src-mediated Tyrosine residues on p97Eps8 and Tyr-45 was one of them, which was not present on p68Eps8. In this study, we utilized Site-directed mutagenesis and Mass spectrophotometer to identify the Src-mediated phosphorylation sites on p68Eps8. We found that both Tyr-524 and Tyr-674 on p68Eps8 were the two preferential phosphorylation residues mediated by Src both in vivo and in vitro.

中文摘要 3
英文摘要 4
縮寫簡表 7
第一章序論 9
第二章實驗材料及方法 15
第一節實驗材料 16
第二節實驗方法 18
第三章實驗結果 34
第四章討論 40
圖表 44
參考文獻 61

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