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Title: | 鯉魚腎泌分解?作用於頭腎細胞間質之生化證據 Biochemical Action of Carp Nephrosin in the Extracellular Matrix of Headkidney |
Authors: | Ching-Yi Chan 詹靜怡 |
Keyword: | 頭腎、腎泌分解?、細胞間質、耦合金屬蛋白?、組織萃取。, headkidney, nephrosin, astacin, extracellular matrix, glycosaminoglycan., |
Publication Year : | 1997 |
Degree: | 碩士 |
Abstract: | 鯉魚頭腎兼具神經、內分泌、免疫、造血組織,後兩者的細胞常有旺盛的細胞遷移活動,作用於細胞外間質的蛋白質水解酵素是媒介此過程重要的分子之一。我們實驗室先前在頭腎純化到一個分泌性的耦合金屬蛋白質水解酵素,由於它主要分佈於頭腎、腎臟及脾臟,而這些器官與造血、免疫功能密切相關,故推測它是作用於細胞外間質。經由多株抗體以免疫篩選得到這個蛋白?的cDNA,由一級結構的保守性,確定它是astacin family的一個新的成員。由其分佈器官特異性,我們將之命名為腎泌分解?(nephrosin)。這個家族成員一般主要功能與胚胎發育、組織重建有關,有許多成員的作用對像是細胞外間質的成分蛋白。因此我們測驗腎泌分解?對幾種哺乳類的細胞外間質組成蛋白質的水解能力,初步結果顯示腎泌分解?會水解fibronectin,它是basement membrane組成之一,牽涉到細胞附著、遷移。隨後我們純化鯉魚的fibronectin,以確定它是頭腎中腎泌分解?的可能天然受質。 腎泌分解?在這個家族中的特殊性在於它mature form的蛋白質結構,只有一個用於水解的proteolytic domain,而該家族的其他成員除此之外大多在proteolytic domain的C-端具有參與蛋白質交互作用的結構,媒介與細胞外間質組成的作用。由於腎泌分解?具有heparin binding的性質,本論文探討它藉proteoglycan上的glycosaminoglycan結合於細胞外間質的可能性。主要的發現是當萃取溶液含glycosaminoglycan類分子時,可以從細胞外間質組成萃取更多腎泌分解?,這表示腎泌分解?可能藉proteoglycan上的glycosaminoglycan結合於細胞外間質。 Our laboratory purified a 23 kDa secreted metalloprotease from carp headkidney, an organ consists of nervous, endocrine, immune, and hematopoietic tissues. The cDNA encoding this protease was isolated and it turned out to be a new member of the astacin family of metalloproteases. This thesis presents the biochemical action of nephrosin in the extracellular matrix of headkidney. Since the cells in hematopoietic tissues are active in migratory events, in which remodeling of extracellular matrix (ECM) usually involves. First, It was examined if nephrosin degraded the common components of ECM. Among the proteins tested, including type IV collagen, fibronectin, and laminin, only bovine fibronectin was degraded by nephrosin. Then carp fibronectin was purified and it was also degraded by nephrosin, raising the possibility that fibronectin is a natural substrate of nephrosin. The mature form of nephrosin is composed of a single proteolytic domain, different from most members of the astacin family, which contain protein-interaction domains C-terminal to their proteolytic domains. Since nephrosin is a heparin binding protein, the hypothesis that nephrosin binds to extracellular matrix via the glycosaminoglycan chain(s) of the proteoglycan(s) was tested. It was found that buffers containing chondroitin sulfate B, dextran sulfate, heparin, heparan sulfate, de-N-sulfated heparin, and high concentrations of NaCl all facilitated the extraction of nephrosin from headkidney ECM-like materials. Therefore, all the results indicate that secreted nephrosin is targeted to ECM by binding to proteoglycans. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76293 |
Fulltext Rights: | 未授權 |
Appears in Collections: | 動物學研究所 |
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