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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 孔繁璐(Fan-Lu Kung) | |
dc.contributor.author | Ke-Ming Huang | en |
dc.contributor.author | 黃可名 | zh_TW |
dc.date.accessioned | 2021-06-15T02:57:08Z | - |
dc.date.available | 2014-10-07 | |
dc.date.copyright | 2011-10-07 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-08-16 | |
dc.identifier.citation | 1. Hardy, J. (1997) Amyloid, the presenilins and Alzheimer's disease, Trends Neurosci 20, 154-159.
2. Hardy, J. A., Mann, D. M., Wester, P., and Winblad, B. (1986) An integrative hypothesis concerning the pathogenesis and progression of Alzheimer's disease, Neurobiol Aging 7, 489-502. 3. Esler, W. P., and Wolfe, M. S. (2001) A portrait of Alzheimer secretases--new features and familiar faces, Science 293, 1449-1454. 4. Corder, E. H., Saunders, A. M., Strittmatter, W. J., Schmechel, D. E., Gaskell, P. C., Small, G. W., Roses, A. D., Haines, J. L., and Pericak-Vance, M. A. (1993) Gene dose of apolipoprotein E type 4 allele and the risk of Alzheimer's disease in late onset families, Science 261, 921-923. 5. Kuo, Y. M., Emmerling, M. R., Bisgaier, C. L., Essenburg, A. D., Lampert, H. C., Drumm, D., and Roher, A. E. (1998) Elevated low-density lipoprotein in Alzheimer's disease correlates with brain abeta 1-42 levels, Biochem Biophys Res Commun 252, 711-715. 6. Jick, H., Zornberg, G. L., Jick, S. S., Seshadri, S., and Drachman, D. A. (2000) Statins and the risk of dementia, Lancet 356, 1627-1631. 7. Katzov, H., Bennet, A. M., Hoglund, K., Wiman, B., Lutjohann, D., Brookes, A. J., Andreasen, N., Blennow, K., De Faire, U., and Prince, J. A. (2006) Quantitative trait loci in ABCA1 modify cerebrospinal fluid amyloid-beta 1-42 and plasma apolipoprotein levels, J Hum Genet 51, 171-179. 8. Refolo, L. M., Malester, B., LaFrancois, J., Bryant-Thomas, T., Wang, R., Tint, G. S., Sambamurti, K., Duff, K., and Pappolla, M. A. (2000) Hypercholesterolemia accelerates the Alzheimer's amyloid pathology in a transgenic mouse model, Neurobiol Dis 7, 321-331. 9. David L. Nelson, M. M. C. (2005) Principles of Biochemistry, 4th ed., W. H. Freeman and Company, New York. 10. Bouillot, C., Prochiantz, A., Rougon, G., and Allinquant, B. (1996) Axonal amyloid precursor protein expressed by neurons in vitro is present in a membrane fraction with caveolae-like properties, J Biol Chem 271, 7640-7644. 11. Riddell, D. R., Christie, G., Hussain, I., and Dingwall, C. (2001) Compartmentalization of beta-secretase (Asp2) into low-buoyant density, noncaveolar lipid rafts, Curr Biol 11, 1288-1293. 12. Wolozin, B. (2001) A fluid connection: cholesterol and Abeta, Proc Natl Acad Sci U S A 98, 5371-5373. 13. Cordy, J. M., Hussain, I., Dingwall, C., Hooper, N. M., and Turner, A. J. (2003) Exclusively targeting beta-secretase to lipid rafts by GPI-anchor addition up-regulates beta-site processing of the amyloid precursor protein, Proc Natl Acad Sci U S A 100, 11735-11740. 14. Epand, R. M. (2006) Cholesterol and the interaction of proteins with membrane domains, Prog Lipid Res 45, 279-294. 15. Beel, A. J., Mobley, C. K., Kim, H. J., Tian, F., Hadziselimovic, A., Jap, B., Prestegard, J. H., and Sanders, C. R. (2008) Structural studies of the transmembrane C-terminal domain of the amyloid precursor protein (APP): does APP function as a cholesterol sensor?, Biochemistry 47, 9428-9446. 16. Kamata, K., Manno, S., Ozaki, M., and Takakuwa, Y. (2008) Functional evidence for presence of lipid rafts in erythrocyte membranes: Gsalpha in rafts is essential for signal transduction, Am J Hematol 83, 371-375. 17. 劉亭儀. (2010) APP與flotillin-1之間的交互作用對APP processing的影響. 18. Beel, A. J., Sakakura, M., Barrett, P. J., and Sanders, C. R. (2010) Direct binding of cholesterol to the amyloid precursor protein: An important interaction in lipid-Alzheimer's disease relationships?, Biochim Biophys Acta 1801, 975-982. 19. Mok, S. S., Sberna, G., Heffernan, D., Cappai, R., Galatis, D., Clarris, H. J., Sawyer, W. H., Beyreuther, K., Masters, C. L., and Small, D. H. (1997) Expression and analysis of heparin-binding regions of the amyloid precursor protein of Alzheimer's disease, FEBS Lett 415, 303-307. 20. Gralle, M., Oliveira, C. L., Guerreiro, L. H., McKinstry, W. J., Galatis, D., Masters, C. L., Cappai, R., Parker, M. W., Ramos, C. H., Torriani, I., and Ferreira, S. T. (2006) Solution conformation and heparin-induced dimerization of the full-length extracellular domain of the human amyloid precursor protein, J Mol Biol 357, 493-508. 21. Chattopadhyay, A., and London, E. (1988) Spectroscopic and ionization properties of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-labeled lipids in model membranes, Biochim Biophys Acta 938, 24-34. 22. Lin, S., and Struve, W. S. (1991) Time-resolved fluorescence of nitrobenzoxadiazole-aminohexanoic acid: effect of intermolecular hydrogen-bonding on non-radiative decay, Photochem Photobiol 54, 361-365. 23. Lee, M. S., Kao, S. C., Lemere, C. A., Xia, W., Tseng, H. C., Zhou, Y., Neve, R., Ahlijanian, M. K., and Tsai, L. H. (2003) APP processing is regulated by cytoplasmic phosphorylation, J Cell Biol 163, 83-95. 24. Koshino, I., and Takakuwa, Y. (2009) Disruption of lipid rafts by lidocaine inhibits erythrocyte invasion by Plasmodium falciparum, Exp Parasitol 123, 381-383. 25. Kamiya, Y., Ohta, K., and Kaneko, Y. (2005) Lidocaine-induced apoptosis and necrosis in U937 cells depending on its dosage, Biomed Res 26, 231-239. 26. Ehehalt, R., Keller, P., Haass, C., Thiele, C., and Simons, K. (2003) Amyloidogenic processing of the Alzheimer beta-amyloid precursor protein depends on lipid rafts, J Cell Biol 160, 113-123. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/44430 | - |
dc.description.abstract | 在阿茲海默氏症(Alzheimer’s disease, AD)的研究領域中,膽固醇與b-amyloid precursor protein (APP)蛋白之間的關係一直是個懸而未決之謎。膽固醇可能借由與APP蛋白有直接的交互作用,而促使APP蛋白走向產生導致阿茲海默氏症的Ab的水解途徑。為了要進一步探討膽固醇與APP蛋白之間的交互作用如何影響APP的水解,我們利用點突變的方式將APP蛋白上與膽固醇做辨認的cholesterol recognition amino acid consensus (CRAC)序列做突變,試著藉由突變產生無法與膽固醇有良好交互作用的APP突變型蛋白。實驗結果發現,APP突變型與lipid raft的標誌蛋白flotillin-1 共分層的程度比野生型來得少;APPY422L與APPS622L在經常用來評估引發致病水解途徑的C99/C83比值上,也較野生型來得低;更進一步的研究發現在去除了膜上的膽固醇後,和僅僅破壞膜上的raft構造,但膜上的膽固醇含量卻仍維持不變時相比,APP蛋白的分層情況並不相同。這些結果指出膽固醇與APP蛋白之間的交互作用力可能造成APP蛋白更容易散布進入lipid raft中,使得APP蛋白更有機會被BACE1水解酶水解,產生Ab片段。 | zh_TW |
dc.description.abstract | The relationship between cholesterol and b-amyloid precursor protein (APP) processing remains an unsolved mystery in the Alzheimer’s disease (AD) research field. It has been hypothesized that there is a direct interaction between cholesterol and APP, which might facilitate APP being processed into Ab in its pathogenic pathway. To assess the importance of the APP-cholesterol in the regulating of regulating APP processing, we generated several cholesterol-binding-deficient APP mutant proteins via introducing mutations in the domains with the putative cholesterol recognition amino acid consensus (CRAC) sequence of APP. Our results revealed that a smaller portion of APP mutants was co-fractionated with the lipid raft marker flotillin-1 compared to wild-type APP. The C99/C83 ratio, which is usually used in evaluating pathogenesis processing level, is also reduced by Y422L and S622L mutations. Further study shows APP pattern in cholesterol-depleted cells was different from raft-disrupted cells in which the membrane cholesterol content remained unchanged. These data suggested that the interaction between cholesterol and APP could be a driving force for distributing APP into lipid raft, which makes APP more accessible to the pathogenically critical protein BACE1. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T02:57:08Z (GMT). No. of bitstreams: 1 ntu-100-R97423028-1.pdf: 2684258 bytes, checksum: 0c13505be8a98ecde8fb9c511add2e12 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 口試委員會審定書…………………………………………………… I
誌謝…………………………………………………………………… II 中文摘要……………………………………………………………… III 英文摘要……………………………………………………………… IV Abbreviations ……………………………………………………… V Contents ……………………………………………………………… VII Introduction …………………………………………………………… 1 Material and Methods ……………………………………………… 4 Site-directed mutagenesis ………………………………………… 4 GST-APP fusion protein purification ………………………… 5 NBD-cholesterol binding assay ………………………………… 6 HEK293T cell transfection ……………………………………… 7 Protein lysate extraction ……………………………………… 8 Western blot ………………………………………………………… 8 Sucrose gradient fractionation ………………………………… 9 Silver stain ………………………………………………………… 9 Cholesterol assay …………………………………………………10 MTT assay ……………………………………………………………… 10 Lipid extraction …………………………………………………… 11 Results ………………………………………………………………… 12 APP mutants …………………………………………………………… 12 NBD-cholesterol binding assay ……………………………………13 Sucrose gradient fractionation ………………………………… 14 C99/C83 ratio …………………………………………………………16 MβCD and lidocaine treatment ……………………………………17 Discussion …………………………………………………………… 19 Cholesterol recognition amino acid consensus (CRAC) and APP mutants …19 NBD-cholesterol binding assay ……………………………………… 21 Sucrose gradient experiment and C99/C83 ratio assay ……… 23 MβCD and lidocaine treatment …………………………………… 24 Figures ……………………………………………………………… 26 Tables ……………………………………………………………… 44 References …………………………………………………………… 45 | |
dc.language.iso | en | |
dc.title | 膽固醇在APP蛋白質水解反應中所扮演的角色 | zh_TW |
dc.title | The Role of Cholesterol in Aβ Precursor Protein Processing | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 李財坤(Tsai-Kun Li),兵岳忻(Yueh-Hsin Ping),顧記華(Jih-Hwa Guh) | |
dc.subject.keyword | 膽固醇,APP蛋白,CRAC序列,lipid raft,C99/C83比值, | zh_TW |
dc.subject.keyword | cholesterol,APP protein,CRAC sequence,lipid raft,C99/C83 ratio, | en |
dc.relation.page | 47 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2011-08-16 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 藥學研究所 | zh_TW |
顯示於系所單位: | 藥學系 |
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