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標題: | 硫酸化酪氨酸探針之設計 Design of Probes for Sulfo-Tyrosine |
作者: | Yen-Min Hsu 許諺銘 |
指導教授: | 吳世雄 |
關鍵字: | 蛋白酪氨酸硫酸化酶,等溫滴定微量熱儀, tyrosylprotein sulfotransferase,Tyrosine O-sulfation,Phos-tag,sulf-tag,nitrilotriacetic acid,Isothermal titration calorimetry, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 蛋白酪氨酸硫酸化是一種轉譯後修飾,此修飾在生理上有重要的功能。但是被發現硫酸化的蛋白量很少。目前磷酸化的酪氨酸已經有商品化的探針可以辨識被,但硫酸化的酪氨酸沒有。因此我們參考可辨識被磷酸化酪氨酸的探針,利用它結合磷酸的策略設計硫酸化酪氨酸的探針,以研究硫酸化蛋白質體。本實驗使用磷酸探針,一個磷酸探針類似物,次氮基三乙酸以及3-((2-氨基苯基)氨基甲醯基)苯甲酸與錳,鈷,鎳,銅,鋅,鈣,鍶及鋇的二價離子形成錯合物,然後測試這些錯合物與苯基磷酸或4-硝基苯基硫酸的結合力。實驗結果顯示,次氮基三乙酸/鍶或鋇可以與4-硝基苯基硫酸有結合能力,且次氮基三乙酸/鋇有較好的選擇性。根據這些結果可知鋇離子具有成為磷酸探針中心離子的潛力。因此,未來將專注在次氮基三乙酸的修飾以增加硫酸化酪氨酸探針的效能。 Protein tyrosine O-sulfation is a post-translational modification (PTM) which was first observed in 1954. Tyrosine-sulfated proteins play the important roles in many biological processes. However, few sulfated proteins have been found by proteomics. Nowadays, protein tyrosine phosphorylation can be recognized by a commercial probe called Phos-tag but no sulf-tag was discovered and applied to sulfoproteomics. Therefore, the specific aim of this study is to design a series of sulf-tags similar to Phos-tag for the enrichment of sulfotyrosine peptides for sulfoproteomics study. We used Phos-tag, a Phos-tag analog, nitrilotriacetic acid and 3-((2-Aminophenyl)carbamoyl)benzoic acid to form complexes with Mn2+, Co2+, Ni2+, Cu2+, Zn2+ ,Ca2+, Sr2+, Ba2+, and then measured the binding ability between complexes and phenyl phosphate and 4-nitrophenyl sulfate respectively. The results showed that nitrilotriacetic acid/Sr2+ and Ba2+ could bind to 4-nitrophenyl sulfate, and nitrilotriacetic acid/Ba2+ had higher selectivity than nitrilotriacetic acid/Sr2+. On the basis of these results, Ba2+ might be the best potential metal ion to build the ideal model for pho-stag. In the future, we will focus on the modification of nitrilotriacetic acid to improve the sulf-tag. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61374 |
全文授權: | 有償授權 |
顯示於系所單位: | 生化科學研究所 |
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