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系統識別號 U0026-0812200910422023
論文名稱(中文) 研究v-Src磷酸化p68Eps8的胺基酸定序
論文名稱(英文) Studies of v-Src-mediated p68EPS8 phosphorylation
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 91
學期 2
出版年 92
研究生(中文) 史披樂
研究生(英文) Pi-Le Shih
電子信箱 overcomerpillar@yahoo.com.tw
學號 s2690411
學位類別 碩士
語文別 中文
論文頁數 68頁
口試委員 指導教授-呂增宏
口試委員-張文昌
口試委員-賴明德
口試委員-馬明琪
中文關鍵字 磷酸化 
英文關鍵字 Eps8  Src 
學科別分類
中文摘要 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。
英文摘要 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.
論文目次 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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