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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18191
標題: | 探索發報基團在化學探針新應用之研究 Manipulations of the Reporter Group for Versatile Applications of Chemical Probes |
作者: | Chih-Hsuan Tai 戴志軒 |
指導教授: | 羅禮強(Lee-Chiang Lo) |
關鍵字: | 活性探針,硫酸酯水解酵素,螢光探針,共軛焦顯微鏡,氟[碳]相催化劑,微波反應,綠色化學,絲胺酸水解型酵素,同位素編碼親和標籤, activity-based probes,sulfatase,fluorogenic probes,confocal fluorescence microscopy,fluorous catalyst,microwave reaction,green chemistry,serine hydrolase,ICAT, |
出版年 : | 2015 |
學位: | 博士 |
摘要: | 活性探針獨特的活性標示能力已被廣泛地應用於蛋白質體學之研究,運用其在生物系統中專一性地反應特定目標蛋白分子的活性,以研究生物系統在特定生理狀態下的酵素活性調控。而本論文的工作致力於化學探針發報端的開發,增加其研究的應用,共進行了三種不同的開發方向,第一個是設計、合成與應用硫酸酯水解酵素活性螢光探針,第二為發展與應用氟[碳]相催化劑,第三為發展具有同位素編碼親和標籤之絲胺酸水解型酵素活性探針。在第一個部分,我們設計一組螢光探針,具備辨識與標記硫酸酯水解酵素的能力,並於標記後開啟螢光訊號,配合共軛焦顯微鏡,可應用於細胞中硫酸酯水解酵素之活性顯影。第二部份的研究為合成與應用氟[碳]相有機金屬與有機催化劑,利用氟[碳]相化合物具備容易純化的特性,發展氟[碳]相催化劑,搭配微波反應進行催化,並且有效率地純化產物與回收催化劑,達到綠色化學的訴求。最後一部分的研究為針對絲胺酸水解型酵素發展具備同位素標籤之活性探針,結合質譜儀系統,增進在複雜系統中酵素活性的定量研究。 The activity-based probes have been widely used in proteomic researches due to its ability to reflect specific enzyme’s activity under certain physiological conditions. We have put a lot of effort into reporter group manipulation for versatile application of chemical probes. We reported three kinds of developments in this dissertation. First, design, synthesis and application of turn-on fluorescent probes for imaging steroid sulfatase activities in cells. Secondly, development and application of fluorous catalysts. Thirdly, development of isotope-coded ABPs for quantitative profiling of serine hydrolase. In the first part, we have designed two turn-on fluorescent probes targeting steroid sulfatase which are designed to generate immobilized fluorescent signal upon hydrolysis. Confocal fluorescence imaging experiments were carried out and compared the results to commercially available imaging reagent. The second part includes development of organo- and organometallic fluorous catalysts. Due to the characters of easy separation, fluorous chemistry is widely utilized in organic synthesis. We have developed new fluorous-taged catalysts and carried catalytic reactions under microwave irradiation. These catalysts did offer better efficiency in purification and matched the statements of green chemistry. The last part is the development of isotope-coded ABPs which applied for quantitative profiling of SHs. We envisioned this design would benefit from MS analysis platform that ensure the in-depth study of complex proteomes. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/18191 |
全文授權: | 未授權 |
顯示於系所單位: | 化學系 |
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ntu-104-1.pdf 目前未授權公開取用 | 9.36 MB | Adobe PDF |
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