白細胞介素6（Interleukin-6；IL-6）是細胞因子的一種，其在人體中主要藉由經典信號傳遞以及反式信號傳遞引發一系列生理反應，廣泛的功能使IL-6無論在抵抗細菌、病毒感染或是在腫瘤中促進腫瘤生成、以及在各種炎症或發炎反應中都扮演重要的角色，尤其反式信號傳遞更被認為是促進腫瘤以及各種自身免疫性炎症病情發展的主因。在之前已有研究報導發現IL-6/sIL-6R的二聚體與gp130之間的鍵結能被肝素抑制，且IL-6與IL-6R之間的結合也能被肝素部分抑制。為了從結構上瞭解IL-6與醣類之間的作用，我們利用圓二色吸收光譜及核磁共振光譜探討IL-6與化學合成硫酸肝素(HS8)之間的作用。並發現IL-6與HS8鍵結時並不會影響IL-6的二級結構，而IL-6主練循序判定完成率為15NH：92%；15N：92%；Cα：92%；Cβ：68%；CO：80%，並初步發現可能會與HS8發生鍵結的正電胺基酸。

Interleukin-6 (IL-6), a member in cytokine family, plays an important role in cell signaling. It plays mainfold function in the processes of bacterial infection resistance, tumor growth, and in various inflammatory diseases. IL-6 initiates these physiological reactions through classical signaling and trans-signaling pathway in human body. Especially, the trans-signaling pathway is considered the main cause in promoting tumors growth and the developing autoimmune inflammatory diseases. Trans-signaling pathway is triggered by the binding of IL-6 to soluble IL-6 receptor (sIL-6R); the IL6/sIL-6R complex then activates the phosphorylation of gp130, and relays the signal to the following members in the cascade. Various inhibitors that regulate the trans-signaling pathway by inhibiting the activation process of gp130 were found. Among these inhibitors, heparin molecules were found to inhibit the interaction of IL-6/sIL-6R complex and gp130. In addition, heparin molecules can partially inhibit the binding between IL-6 and IL-6R. In order to further understand the inhibition process, we used circular dichroism (CD) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy to study the interaction of IL-6 with a model heparin molecule, a chemically synthetic heparin sulfate octasaccharide (HS8). We found that the binding of HS8 onto IL-6 will not change the secondary structure percentage of IL-6.The completeness of backbone assignments of IL-6 is 92%(NH, N, Cα), 68% (Cβ) , 80%(CO).The possible HS binding surface, including R31, R41, K42, R180 and R183 residues is located on same side of IL-6.

一、文獻探討.......1
1.1 白細胞介素6（IL-6）......1
1.2 醣胺聚醣 （Glycosaminoglycan；GAGs）.....4
1.3 蛋白質與硫酸肝素的作用.....6
1.4 研究目的.....8
二、實驗材料與方法.....9
2.1 蛋白質表現.....9
2.2 蛋白質分離、折疊與純化.....10
2.3 圓二色吸收光譜實驗.....11
2.4 DLS（Dynamic light scattering）實驗.....11
2.5 NMR光譜實驗.....12
2.5.1 IL-6的NMR樣品配置.....12
2.5.2 IL-6主鍊的化學位移和循序判定.....13
2.5.3 IL-6與不同濃度HS8滴定實驗.....13
三、實驗結果 .....14
3.1 IL-6蛋白質純化與鑑定.....14
3.2 圓二色吸收光譜實驗.....14
3.3 DLS （Dynamic light scattering）分析IL-6粒徑大小.....15
3.4 IL-6蛋白質的NMR光譜條件實驗.....16
3.5 IL-6主鍊循序判定.....17
3.6 IL-6與HS8滴定實驗.....18
四、討論.....19
五、參考文獻.....23

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