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研究生:黎正源
研究生(外文):Zheng-Yuan Lee
論文名稱:探討C反應蛋白的結構與功能的關係
論文名稱(外文):Structure and Function Study of C-reactive protein
指導教授:許先業
指導教授(外文):Hsien-Yeh Hsu
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:醫學生物技術研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:英文
論文頁數:69
中文關鍵詞:急性期蛋白發炎急性心肌梗塞離子交換原子力顯微鏡
外文關鍵詞:acute phase proteininflammationacute myocardio infractionion exchangeatomic force microscope
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  • 被引用被引用:1
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C反應蛋白是一種屬於五元體環形結構的急性反應期蛋白,最近的報告指出C反應蛋白會引起血管細胞的發炎反應。在這篇論文中,我們的研究針對是否會經由結構的改變而造成促進發炎的反應。首先我們利用冷光光譜儀紀錄到在C反應蛋白變為修飾的C反應蛋白的過程中,內生性的冷光會降,同時圓極偏光光譜計的測量也顯示兩者的二級結構有不同。接著利用階梯式的陰離子交換樹酯的技術將C反應蛋白與修飾的C反應蛋白成功的分離。得到了純的蛋白質後,我們利用原子力顯微鏡及分析級超高速離心機證明了修飾的C反應蛋白是失去五元體環形結構的單體。接著我們顯示C反應蛋白會接合到程式死亡及壞死的細胞上。並且會降低程式死亡細胞及吞噬細胞所產生的活性氧。另一方面,修飾的C反應蛋白會因起THP-1單合球細胞分泌間白素8及間質金屬蛋白脢9。因此,這些結果指出C反應蛋白的結構改變會影響到它的功能性。
Native pentameric C-reactive protein (pCRP), an acute phase protein, belongs to pentraxin family and recently has been suggested to have pro-inflammation effects on vascular cells. In this study, we tried to find out whether conformational changes of pCRP involved in these effects. Using luminescence spectrometer and circular dichroism spectrometer, we recorded the intrinsic decrease of emission fluorescence during the conversion from pCRP to modified CRP (mCRP) and the difference in secondary structure, respectively. A stepwise anion-exchange was used for separating and purifying mCRP. After getting pure pCRP and mCRP, we used atomic force microscopy and analytical ultracentrifugation assays to reveal that mCRP was a dissociated monomer-like, which loss pentameric symmetry. Next, we demonstrated that pCRP binds to apoptotic and necrotic cells and reactive oxygen species was attenuated in pCRP-bound apoptotic cells as well as phagocytes internalizing them. In the other hand, incubation with mCRP increased IL-8, one of key regulator of monocytes recruitment, and MMP-9, one of molecules responsible for plaque rupture, secretion in THP-1 monocytes. Together, these results establish that changes in structure of pCRP affect its functions.
1. Tillett W. S., Francis T. (1930) Serological reactions in pneumonia with a non-protein somatic fraction of pneumococcus. J. Exp. Med. 52, 561-571.
2. Shine B., De Beer F. C., Pepys M. B. (1981) Solid phase radioimmunoassays for C-reactive protein. Clin. Chim. Acta. 117, 13–23.
3. Gabay C., Kushner I. (1999) Acute-phase proteins and other systemic responses to inflammation. N. Engl. J. Med. 340, 448- 454.
4. Vigushin D. M., Pepys M. B., Hawkins P. N. (1993) Metabolic and scintigraphic studies of radioiodinated human C-reactive protein in health and disease. J. Clin. Invest. 91, 1351–1357.
5. Thompson D., Pepys M. B., Wood S. P. (1999) The physiological structure of human C-reactive protein and its complex with phosphocholine. Structure. 7, 169-177.
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