臺灣博碩士論文加值系統

肝素為目前廣泛被認知的具有多樣生理活性的碳水化合物，它能辨認各式各樣的相關蛋白質並介入其蛋白質-蛋白質間的作用，其中包含了趨化因子( Chemokines ) 以及生長因子 ( Growth factor ) 等。肝癌衍生生長因子被認為是肝癌發展的重要蛋白質，具有促進細胞增殖的功能，在細胞內可與細胞核中的DNA鍵結並作用，於細胞外作為趨化生長因子，並且與細胞膜上的醣類作用，因此，我們利用核磁共振儀器加以探討肝癌生長子因子與肝素類似物 ( 8 mer ) 作用之結構在蛋白質以及醣，以及其各別的結構變化，藉此提出蛋白質-醣作用的結構模型。

Heparan sulfate is a widely recognized carbohydrate with a variety of physiological activities that recognizes a wide variety of related proteins and intervenes in its protein-protein interactions, including chemokines and growth factors, etc. Hepatoma-derived growth factor is considered to be an important protein for the development of hepatocellular carcinoma. It has the function of promoting cell proliferation, which can bind to the DNA in the nucleus and act as a chemotactic growth factor and cell membrane , Therefore, we use nuclear magnetic resonance instruments to explore the role of hepatoma-derived growth factor and heparan sulfate analogue (8 mer) in the structure of proteins and sugars, as well as their respective structural changes, whereby the proposed structural model demonstrates .

目錄
一、 文獻探討
1.1 肝癌 1
1.2 肝癌衍生生長因子 1
1.3 糖胺聚醣 ( Glycosaminoglycan ) 3
1.4 蛋白質與醣的作用模型 5
1.5論文研究目的 6
二、 實驗材料與方法
2.1 蛋白質的表現 7
2.2 蛋白質純化 7
2.3 HDGF-N100之NMR光譜實驗與分析 8
2.4 HDGF-N100主鏈的化學位移循序判定 8
2.4.1 HNCA、HN(CO)CA 8
2.4.2 HNCO、HN(CA)CO 8
2.4.3 CBCANH、CBCA(CO)NH 9
2.5 HDGF-N100側鏈的化學位移判定 9
2.5.1 HCCH-COSY、HCCH-TOCSY 9
2.6 HS8 的 NMR 光譜實驗與分析 9
2.7 HS8化學位移判定 10
2.7.1 HS8 耦合常數與二面角判定 10
2.7.2 NOESY-HSQC的分子間化學位移判定，分子內( Intra )、分子間
( Inter )距離計算 10
2.8 鍵結的HDGF-N100的結構計算 11
2.9 鍵結的HS8的結構計算 12
2.10 HDGF-N100、HS8作用後的結構計算 12
三、 實驗結果與討論
3.1 蛋白質的純化與鑑定 13
3.2 HDGF-N100 主鏈之循序判定 13
3.2.1鍵結配體 ( Free form ) HDGF-N100化學位移 13
3.2.2 HDGF-N100與HS8鍵結 ( Bound form ) 後的化學位移判定 14
3.2.3 HDGF的自由/鍵結滴定實驗比例的校正 15
3.2.4化學位移偏差與解離常數 16
3.3 HDGF-N100之二級結構預測 17
3.4 HDGF-N100結構計算 17
3.5 HS8 自由/鍵結的化學位移判定 18
3.6 HS8耦合常數、距離分析 19
3.7 鍵結的HS8的結構計算 19
3.8 HDGF-HS8 的錯合物的模擬結構 20
3.9 結論 20
四、參考文獻 21
表目錄
圖目錄

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