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標題: | 針對蛋白質酪胺酸磷酸酯水解酵素開發具有不同發報端的活性標示分子 Synthesis of activity-based probes for protein tyrosine phosphatases with various reporter groups for versatile applications |
作者: | Chao-Kai Chen 陳昭凱 |
指導教授: | 羅禮強(Lee-Chiang Lo) |
關鍵字: | 蛋白質酪胺酸磷酸酯水解酵素,活性標示分子,生物素,螢光團,疊氮基,雙硫鍵, Protein tyrosine phosphatases (PTPs),activity-based probes,2-fluoromethylphenoxyl group,biotin,fluorophore,azido group,disulfide, |
出版年 : | 2009 |
學位: | 碩士 |
摘要: | 目前已知蛋白質酪胺酸磷酸酯水解酵素(Protein Tyrosine Phosphatases, PTPs)參與調控許多細胞間訊號傳導的速率和過程,PTPs的活性表現與許多疾病間都具有關聯性,如癌症、神經失調和糖尿病等。本篇論文針對PTPs發展不同用途的活性標示分子來更加了解它在生物體內特定的動態表現。我們設計並合成出四個活性標示分子,具備了相同的磷酸化酪胺酸類似物作為對PTPs有專一性的辨識端,和連接橋連接潛在的捕捉基團—2-氟甲基-苯氧基基團和不同的發報端或親和標記用來觀察標示結果或純化蛋白質。首先,由酪胺酸開始建構對PTPs的辨識端,接著利用三甘醇衍生得到一端為疊氮基或帶有保護基的胺基作為連接橋,再與辨識端藕合得到具有疊氮基的活性標示分子前驅物,和其他活性標示分子的共同中間體,進一步連接上生物素、螢光團及具雙硫鍵的發報端,最後去除所有磷酸根的丙烯基保護基即完成四個活性標示分子的建構。未來將陸續完成這一系列活性標示分子的標示實驗並進一步和過去的成果比較及評估其發展潛力。 A number of protein tyrosine phosphatases (PTPs) are linked to diseases, ranging from cancers to neurological disorders and diabetes. It was believed that PTPs were involved in controlling the rate and duration of the signal response. The main goal in this thesis is developing different uses of activity-based probes for profiling PTPs in dynamic systems. We designed and synthesized four activity-based probes that carry the same phosphotyrosine analog as a PTP-specific recognition head and a linker that connects the latent trapping device, 2-fluoromethylphenoxyl group, with different reporter groups or affinity tags for visualization and purification. First, a recognition head for PTPs was constructed from tyrosine and linkers with one side carrying an azido group or protected amino group were derived from triethylene glycol. Afterward a precursor of azido-based probe and common intermediates of probes were obtained by coupling the recognition head and linkers. Further, connecting reporter groups of biotin, fluorophore and disulfide-containing linker to the intermediates, and then removing all the allyl protecting groups of phosphates to afford the four probes. All the labeling performance and versatile applications for these probes will be compared to our previous work and evaluated in the future. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26107 |
全文授權: | 未授權 |
顯示於系所單位: | 化學系 |
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