2014-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/643234摘要：目前結合化學分析的方式已被廣泛應用在生物系統上，例如酵素抑制劑/激活劑，酵素受質，分子探針及蛋白質修飾劑。透過設計化合物除了能幫助我們掌控蛋白質的功能及蛋白質被修飾的過程外，亦可用來釐清過去傳統生化或遺傳技術在生物層面上所無法解決的問題。先前我們以酵素作用機制為基礎，已成功地設計出一系列可專一性標定水解酶像是酪胺酸磷酸酶和糖苷酶的化學活性探針。在這裡，我們擬進一步利用化學探針標定水解酶的特性，去探討絲胺酸水解酶在大腸癌轉移和侵襲的過程中所扮演的角色。絲氨酸水解酶家族的成員參與細胞中許多重要的生理過程，包括細胞生長，組織分化，腫瘤形成，以及腫瘤的轉移。然而，由於過去缺乏合適的分析方式能夠觀察這些生理過程，因此目前對於了解絲氨酸水解酶如何參與在細胞中的訊息仍然有限。為了解決這個問題，我們將設計並合成一系列針對絲氨酸水解酶具有專一性的化學活性探針，進而分析絲氨酸水解酶在細胞生理過程中所扮演的角色。我們期待在蛋白質體學的研究方法上，這些探針可以提供一個良好的研究平台來分析絲氨酸水解酶的活性並了解其功能，藉此找到與疾病相關的重要標的。本研究的最終目標是想找出有哪些絲氨酸水解酶會參與在疾病的過程中及其所扮演的角色。現階段本研究的目的是藉由開發出具有專一性標定水解酶這類重要家族蛋白的化學活性探針，來分析在癌細胞轉移及上皮至間質轉變過程中絲氨酸水解酶的活性（子計畫2 和3），進而作為檢測結腸癌的工具之一（子計畫3）。此外，我們也設計探針用來觀察絲氨酸水解酶的活性及其對受質的選擇性。本子計畫的具體目標：1、設計並合成對絲氨酸水解酶具有選擇性接合能力的化學活性探針。2、建立經由結合具選擇性的化學活性探針及其他分析方式的研究平台來探討絲氨酸水解酶在生物系統中所扮演的角色。<br> Abstract: Chemical approaches have been used in analyzing biological systems such as enzymeinhibitors/activators, substrates, molecular probes, and protein modifiers. With properdesigned chemical compounds, the function of proteins could be controlled, modified,monitored, or manipulated. This chemistry-driven approach can be used to answer biologicalquestions that cannot be easily addressed by traditional biochemical or genetic approaches.Previously, we have adopted a mechanism-based approach and developed activity probes forhydrolases such as tyrosine phosphatases and glycosidases. We have been successfullydesigned and synthesized a series of activity probes to characterize the activity of these twofamilies of hydrolases. Here we are interested in identifying and analyzing the roles of serinehydrolases during the invasion and metastasis of colorectal cancer using chemical approaches.Members of serine hydrolyase family are crucial for the regulation of numerous cellularevents, including cell growth, tissue differentiation, neoplastic formation, as well as tumormetastasis. However, due to the lack of a proper tool that enables the analysis of them, thecurrent characterization of serine hydrolases preceded at a slower pace. We propose to extendour efforts to design and synthesize chemical probes for serine hydrolases analyses. Ourapproach is unique in that the designs of probes could only target to serine hydrolases. Weanticipate that our chemical approach will provide an excellent technical platform inproteomic research, serine hydrolases activity analysis, and facilitate the process in findingimportant disease-related targets. Moreover, these activity probes will provide critical analysistools that will greatly facilitate our understanding of serine hydrolases.The long-term objectives are to profile and identify specific serine hydrolases that playimportant roles in disease-related processes. The goal of this proposed research is to developchemical probes that could specifically label to this physiologically important enzyme family.The probes prepared in this study will be used to profile and identify serine hydrolases inmetastasis and epithelial-to-mesenchymal transition (sub-project 2 and 3) and as a tool indetecting markers for colon cancer (Sub-project 3). Moreover, these probes will also be usedas a tool in determine the activity and substrate specificity of serine hydrolases.Specific aims of this sub-project are:1. To design, synthesize, and evaluate novel chemical structures of the trapping device forthe development and evaluation of activity probes with tunable specificity.2. To establish technical platforms with the use of class-selective probes in conjunction withother analyses to study the role of serine hydrolases in biological systems.