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研究生:梁筑婷
研究生(外文):Chu-Ting Liang
論文名稱:利用核磁共振光譜學標定人類脂蛋白元E4氮-端之骨架原子化學位移
論文名稱(外文):Backbone Resonance Assignment of Apolipoprotein E4 N-terminal Domain
指導教授:林達顯
指導教授(外文):Ta-Hsien Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生命科學暨基因體科學研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:77
中文關鍵詞:人類脂蛋白元E阿茲海默氏症
外文關鍵詞:apolipoprotein EAlzheimer’s disease
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人類脂蛋白元E (apolipoprotein E, ApoE)是一種脂質運輸蛋白,由299個胺基酸所組成,分子量大約為34 kDa,它在脂質代謝中扮演非常重要的角色。APOE基因主要有三種對偶基因,分別是ε2、ε3、ε4,決定了三種主要的異構物(isoforms) ApoE2、ApoE3及ApoE4。三種異構物兩兩之間的差異只有一個胺基酸不同,位在殘基112或158。ApoE藉著一段鉸鏈區域(hinge region)分成兩個結構區段,氮-端區段(N-terminal domain)由殘基1-191所組成,其功能為負責與低密度脂蛋白接受器結合,碳-端區段(C-terminal domain)則是由殘基216-299所組成,此區段含有與脂蛋白結合的區域。ApoE對脂蛋白的結合程度因異構物種類的不同而有所差異。ApoE4被認為是造成阿茲海默症以及粥狀動脈硬化的主要危險因子。ApoE三種異構物雖只有一個胺基酸差異,但其功能特性卻大不相同,研究認為這可能是由於不同的ApoE異構物其氮-端與碳-端區段間之交互作用(N- and C-terminal domain interaction)有所不同。目前對於此交互作用的分子機制仍不甚明瞭,全長ApoE異構物的結構資訊可幫助我們從結構的觀點瞭解此交互作用之分子機制。目前尚無全長ApoE異構物的三級結構資訊,只有氮-端區段在無脂質條件下之三級結構資訊,因此我們最終之目的是獲得全長ApoE異構物的三級結構資訊,藉此幫助我們瞭解ApoE異構物氮-端與碳-端區段間交互作用之分子機制。由於ApoE會自我聚集(self-association),以現今之結構生物學技術,從全長ApoE異構物的結構直接找出參與氮-端與碳-端區段間交互作用的殘基,有其困難度。因此我們的做法是將ApoE異構物分成氮-端與碳-端區段,再利用核磁共振光譜學(NMR spectroscopy)之技術找出參與氮-端與碳-端區段間交互作用的殘基。為達此目的,本研究將先著重在ApoE4氮-端區段骨架原子化學位移之標定。
Human apolipoprotein E (apoE) is a 299-residues lipid-transport protein with a molecular mass of ~34 kDa. It plays an important role in lipid metabolism. APOE gene has three major alleles, ε2, ε3 and ε4, which encode three major isoforms, apoE2, apoE3, and apoE4. The differences between the three isoforms are residues 112 and 158. ApoE contains two independently folded domains (22 kDa N-terminal domain, residues 1-191, and 10 kDa C-terminal domain, residues 216-299) that are separated by a hinge region. The structure of N-terminal domain in the lipid-free state has been determined by x-ray crystallography, which revealed an anti-paralleled four-helix bundle. This domain is responsible for low density liporpteoin (LDL) receptor binding. The structure of C-terminal domain has not been solved yet. This domain contains lipid-binding region. ApoE isoforms show differential preference for lipoprotein particles. ApoE4 is a major risk factor for Alzheimer’s disease and atherosclerosis. The isoform-specific functions of apoE can be ascribed to the isoform-specific domain interactions. So far, the molecular mechanism of apoE domain interactions is not yet clear. Our ultimate goal is to understand the molecular mechanism of ApoE domain interactions from the structural point of view. Structural information of intact apoE isoforms may help us gain more insight into the molecular mechanism of apoE domain interactions. The structural properties of intact ApoE isoforms are largely unknown. Previous studies suggested that C-terminal domain is responsible for tetramerization of ApoE. This property might have hindered the structural determination of intact ApoE isoforms by current structural biology techniques. Thus, our approach is to divide the intact ApoE into N-terminal and C-terminal domains and apply nuclear magnetic resonance (NMR) spectroscopy to identify the residues involved in domain interactions. In order to accomplish the goal, in this study we focused on the backbone resonance assignment of ApoE4 N-terminal Domain.
中文摘要...................................................1
英文摘要...................................................2
致謝.......................................................3
目錄.......................................................4
表目錄.....................................................6
圖目錄.....................................................7
第一章、緒論...............................................9
1.1 脂蛋白元 (Apolipoprotein) .............................9
1.2 人類脂蛋白元E (Human apolipoprotein E) ...............10
1.2.1 Apolipoprotein E之功能特性.........................10
1.2.2 Apolipoprotein E之結構特性.........................10
1.2.3 Apolipoprotein E之氮-端與碳-端區段間之交互作用.....11
1.2.4 Apolipoprotein E之相關疾病.........................12
1.3 研究目的與設計........................................13
第二章、研究材料與方法....................................15
2.1 實驗材料..............................................15
2.1.1 藥品試劑...........................................15
2.1.2 培養液以及溶液.....................................15
2.2 蛋白質樣品之製備......................................16
2.2.1 表達同位素標記之ApoE4(1-191) ......................16
2.2.2 純化同位素標記之ApoE4(1-191) ......................16
2.3 核磁共振實驗..........................................17
2.3.1 核磁共振實驗樣品之製備與實驗條件...................17
2.3.2 核磁共振實驗數據處理...............................18
2.3.3 蛋白質骨架原子化學位移量測.........................18
2.3.4 脂蛋白元E二級結構之判定............................18
第三章、結果..............................................20
3.1 ApoE4(1-191)之表達....................................20
3.2 ApoE4(1-191)之純化....................................20
3.3 核磁共振光譜分析......................................21
第四章、討論..............................................23
文獻參考..................................................24
1. Wasan, K. M., Brocks, D. R., Lee, S. D., Sachs-Barrable, K., and Thornton, S. J. (2008) Nat Rev Drug Discov 7, 84-99
2. Hatters, D. M., Peters-Libeu, C. A., and Weisgraber, K. H. (2006 ) Trends Biochem Sci. 8, 445-454
3. Elshourbagy, N. A., Liao, W. S., Mahley, R. W., and Taylor, J. M. (1985) Proc Natl Acad Sci U S A 82, 203-207
4. Xu, Q., Bernardo, A., Walker, D., Kanegawa, T., Mahley, R. W., and Huang, Y. (2006) J Neurosci 26, 4985-4994
5. Mahley, R. W., and S. C. Rall, J. (2000) Annu Rev Genomics Hum Genet 1, 507-537
6. Mahley, R. W., Weisgraber, K. H., and Huang, Y. (2008) J Lipid Res
7. Drury, J., and Narayanaswami, V. (2005) J Biol Chem 280, 14605-14610
8. Dong, L. M., Wilson, C., Wardell, M. R., Simmons, T., Mahley, R. W., Weisgraber, K. H., and Agard, D. A. (1994) J Biol Chem 269, 22358-22365
9. Dong, L. M., Parkin, S., Trakhanov, S. D., Rupp, B., Simmons, T., Arnold, K. S., Newhouse, Y. M., Innerarity, T. L., and Weisgraber, K. H. (1996) Nat. Struct. Biol. 3, 718-722
10. Luc, G., Bard, J. M., Arveiler, D., Evans, A., Cambou, J. P., Bingham, A., Amouyel, P., Schaffer, P., Ruidavets, J. B., Cambien, F., and et al. (1994) Arterioscler. Thromb. 14, 1412-1419
11. Saunders, A. M., Strittmatter, W. J., Schmechel, D., George-Hyslop, P. H., Pericak-Vance, M. A., Joo, S. H., Rosi, B. L., Gusella, J. F., Crapper-MacLachlan, D. R., Alberts, M. J., and et al. (1993) Neurology 43, 1467-1472
12. Wetterau, J. R., Aggerbeck, L. P., Rall, S. C., Jr., and Weisgraber, K. H. (1988) J Biol Chem 263, 6240-6248
13. Wilson, C., Wardell, M. R., Weisgraber, K. H., Mahley, R. W., and Agard, D. A. (1991) Science 252, 1817-1822
14. Xu, C., Sivashanmugam, A., Hoyt, D., and Wang, J. (2005) J. Biomol. NMR 32, 177
15. Innerarity, T. L., Friedlander, E. J., Rall, S. C., Jr., Weisgraber, K. H., and Mahley, R. W. (1983) J Biol Chem 258, 12341-12347
16. Weisgraber, K. H. (1994) Adv. Protein Chem. 45, 249-302
17. Weisgraber, K. H. (1990) J Lipid Res 31, 1503-1511
18. Segrest, J. P., Jones, M. K., De Loof, H., Brouillette, C. G., Venkatachalapathi, Y. V., and Anantharamaiah, G. M. (1992) J Lipid Res. 33, 141-166
19. Hatters, D. M., Budamagunta, M. S., Voss, J. C., and Weisgraber, K. H. (2005) J Biol Chem 280, 34288-34295
20. Dong, L. M., and Weisgraber, K. H. (1996) J Biol Chem 271, 19053-19057
21. Mahley, R. W., Huang, Y., and Rall, S. C., Jr. (1999) J Lipid Res 40, 1933-1949
22. Schneider, W. J., Kovanen, P. T., Brown, M. S., Goldstein, J. L., Utermann, G., Weber, W., Havel, R. J., Kotite, L., Kane, J. P., Innerarity, T. L., and Mahley, R. W. (1981) J. Clin. Invest. 68, 1075-1085
23. Davignon, J., Gregg, R. E., and Sing, C. F. (1988) Arteriosclerosis 8, 1-21
24. Selkoe, D. J. (1991) Neuron 6, 487-498
25. Huang, Y., Liu, X. Q., Wyss-Coray, T., Brecht, W. J., Sanan, D. A., and Mahley, R. W. (2001) Proc Natl Acad Sci U S A 98, 8838-8843
26. Carter, D. B., Dunn, E., McKinley, D. D., Stratman, N. C., Boyle, T. P., Kuiper, S. L., Oostveen, J. A., Weaver, R. J., Boller, J. A., and Gurney, M. E. (2001) Ann. Neurol. 50, 468-475
27. Haas, C., Cazorla, P., Miguel, C. D., Valdivieso, F., and Vazquez, J. (1997) Biochem J 325 ( Pt 1), 169-175
28. Thal, D. R., Capetillo-Zarate, E., Schultz, C., Rub, U., Saido, T. C., Yamaguchi, H., Haass, C., Griffin, W. S., Del Tredici, K., Braak, H., and Ghebremedhin, E. (2005) Acta Neuropathol 110, 459-471
29. Shobab, L. A., Hsiung, G. Y., and Feldman, H. H. (2005) Lancet Neurol 4, 841-852
30. Bales, K. R., Dodart, J. C., DeMattos, R. B., Holtzman, D. M., and Paul, S. M. (2002) Mol Interv 2, 363-375, 339
31. Kang, D. E., Pietrzik, C. U., Baum, L., Chevallier, N., Merriam, D. E., Kounnas, M. Z., Wagner, S. L., Troncoso, J. C., Kawas, C. H., Katzman, R., and Koo, E. H. (2000) J. Clin. Invest. 106, 1159-1166
32. Andersen, O. M., and Willnow, T. E. (2006) Trends in Neurosciences 29, 687-694
33. Russo, C., Angelini, G., Dapino, D., Piccini, A., Piombo, G., Schettini, G., Chen, S., Teller, J. K., Zaccheo, D., Gambetti, P., and Tabaton, M. (1998) Proc Natl Acad Sci U S A 95, 15598-15602
34. LaDu, M. J., Pederson, T. M., Frail, D. E., Reardon, C. A., Getz, G. S., and Falduto, M. T. (1995) J Biol Chem 270, 9039-9042
35. Ye, S., Huang, Y., Mullendorff, K., Dong, L., Giedt, G., Meng, E. C., Cohen, F. E., Kuntz, I. D., Weisgraber, K. H., and Mahley, R. W. (2005) Proc Natl Acad Sci U S A 102, 18700-18705
36. Wishart, D. S., Bigam, C. G., Yao, J., Abildgaard, F., Dyson, H. J., Oldfield, E., Markley, J. L., and Sykes, B. D. (1995) J. Biomol. NMR 6, 135-140
37. Wishart, D. S., and Sykes, B. D. (1994) J. Biomol. NMR 4, 171-180
38. Wishart, D. S., and Sykes, B. D. (1994) Methods Enzymol. 239, 363-392
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