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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 莊榮輝(Rong-Huay Juang) | |
| dc.contributor.author | Poi-Wei Chuang | en |
| dc.contributor.author | 莊博為 | zh_TW |
| dc.date.accessioned | 2021-06-13T08:08:51Z | - |
| dc.date.available | 2005-07-21 | |
| dc.date.copyright | 2005-07-21 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-21 | |
| dc.identifier.citation | 吳建興 (2000) 裂殖性酵母菌中植物螯合素合成的分離與性質研究。博士論
文。國立台灣大學農業化學系，台北。 吳裕仁 (1995) 酵母菌中鎘結合蛋白質的純化與性質分析。碩士論文。國立台 灣大學農業化學系，台北。 車玉春 (1997) 酵母菌中鎘結合蛋白質 (W) 的純化與性質分析。碩士論文。 國立台灣大學農業化學系，台北。 簡至德 (1998) 裂殖性酵母菌中鎘結合蛋白質的純化與性質分析。碩士論文。 國立台灣大學農業化學系，台北。 Bizily SP, Rugh CL, Meagher RB (2000) Phytodetoxification of hazardous organomercurials by genetically engineered plants. Nat Biotechnol 18: 213-217 Bodenstein-Lang, J., Buch, A., Follmann, H. (1989) Animal and plant mitochondria contain specific thioredoxins, FEBS Lett. 258: 22-26 Cherian MG, Apostolova MD (2000) Nuclear localization of metallothionein during cell proliferation and differentiation. Cell Mol Biol 46 (2): 347-356 Clemens S, Schroeder JI, Degenkolb T (2001) Caenorhabditis elegans expresses a functional phytochelatin synthase. Eur J Biochem 268: 3640-3643 Corbett AH, Silver PA (1996) The NTF2 gene encodes an essential, highly conserved protein that functions in nuclear transport in vivo, J. Biol. Chem. 271: 18477–18484 Dameron CT, Smith BR, Winge DR (1989) Gluthatione-coated cadmium-sulfide crystsllites in Candida glabrata. J. Biol. Chem. 264: 17355-17360 Dameron CT, Smith BR, Winge DR (1989) Gluthatione-coated cadmium-sulfide crystsllites in Candida glabrata. J. Biol. Chem. 264: 17355-17360 F. Allain et al., (2002) Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T lymphocytes to extracellular matrix. Proc. Natl. Acad. Sci. U. S. A. 99: 2714–2719 F. Allain et al., (2002), Interaction with glycosaminoglycans is required for cyclophilin B to trigger integrin-mediated adhesion of peripheral blood T 107 lymphocytes to extracellular matrix. Proc. Natl. Acad. Sci. U. S. A. 99: 2714–2719 Fisher, G, Wittman. Liebold, B Lang, K, Kiefhaber, T, Schmid, F (1999) Nature 337: 476-478 Furey WF, Robbins AH, Clancy LL, Winge DR, Wang BC, Stout CD (1986) Crystal structure of Cd,Zn metallothionein. Science 231: 704-710 Gadd GM (1990) Metal tolerance, in: “Microbiology of extreme environments” .(C., Edwards) : 178-210.Open University Press, Cambridge Gill E, Thummann J, Winnacker E-L, Zenk MH (1988) Induce of heavy metal binding phytochelatins by inoculation of cell cultures in standard media. Plant Cell Rep 7: 375-378 Grill E, Grkeier W, Winnacker E-L, Zenk MH (1985) Phytochelatins: the principal heavy meatl-complexing peptides of higher plants. Science 230: 674-676 Hall, J. L (2002) cellular mechanisms for heavy metal detoxification and tolerance. J Exp. Bot. 53: 1-11 Howden R, Andersen CR, Goldsbrough PB, Cobbett CS (1995a) A cadmiumsensitive, glutathione-deficient mutant of Arabidopsis thaliana. Plant Physiol 107: 1067-1073 Irene Garcia-Higuera , Chrysanthe Gaitatzes§, Temple F. Smith§¶, d Eva J. Neer (1998) Folding a WD Repeat Propeller ROLE OF HIGHLY CONSERVED ASPARTIC ACID RESIDUES IN THE G PROTEIN SUBUNIT AND Sec13* J Biol Chem, Vol. 273, Issue 15: 9041-9049 Jackson PJ, Unkefer CJ, Doolen JA, Watt K, Robinssion NJ (1987) Poly glycine : its role in cadmium resistance in plant cells. Proc Natl Acad Sci USA 84: 6619-6623 K.N. Brazin et al., (2002), Regulation of the tyrosine kinase Itk by the peptidyl-prolyl isomerase cyclophilin A. Proc. Natl. Acad. Sci. U. S. A. 99: 1899–1904 Kägi JH (1991) Overview of metallothionein. Methods Enzymol 205: 613-626 Karin M (1985) Metallothioneins: proteins in search of function. Cell 41: 9-10 Kondo N, Isobe M, Imai K, Goto T (1983) Structure of cadystin, the unit-peptide of cadmium-binding pepttides induced in a fission yeast, Schizosaccharomyces pombe. Tetrahedron Lett 24: 925-928 Laloi C., Rayapurmi N., Chartier Y., Grienenberger J., Bonnard G., Meyer Y (2001) Identification and characterization of a mitochondrial thioredoxin system in plants Proc. Natl. Acad. Sci. U. S. A. 98: 14144-14149 Lees E (1995) Cyclin dependent kinase regulation. Current Opinion in Cell Biology 7: 773–780 Liang ZY, Pilon-Smits EA, Jouanin L, Terry N (1999) Overexpression of 108 glutathione synthetase in indian mustard enhances cadmium accumulation and tolerance. Plant Physiol 199: 73-80 M.Y. Galperin, A. Bairoch, E.V. Koonin (1998) A superfamily of metalloenzymes unifies phosphopentomutase and cofactor-independent phosphoglycerate mutase with alkaline phosphatases and sulfatases. Protein Sci. 7: 1829–1835 Maret W (1994) Oxidative metal release from metallothionein via zinc-thiol/disulfide interchange. Proc Natl Acad Sci U S A 91: 237-241 Mendum ML, Gupta SC, Goldsbrough PB (1990) Effect of glutathione on phytochelatin synthesis in tomato cells. Plant Physiol 93: 484-488 Moffat AS, (1995) Plants proving their worth in toxic metal cleanup. Science 269: 302-303 Myung JK, Gulesserian T, Fountoulakis M, Lubec G (2003) Deranged hypothetical proteins Rik protein, Nit protein 2 and mitochondrial inner membrane protein, Mitofilin, in fetal Down syndrome brain. Cell Mol Biol (Noisy-le-grand).49(5):739-46. Nies, D.H (1999) Microbial heavy–metal resistance. Appl. Microbiol. Biotechnol. 51: 730-570 Pedrajas, J. R., Kosmidou, E., Miranda-Vizuete, A., Gustafsson, J. A., Wright, A. P, Spyrou, G (1999) Identification and Functional Characterization of a Novel Mitochondrial Thioredoxin System in Saccharomyces cerevisiae. J. Biol. Chem. 274: 6366-6373 Rauser WE (1991) Cadmium-binding peptides from plants. Methods Enzymol 205: 319-333 Reese RN, Mehra RK, Tarbet EB, Winge DR (1988) Studies on the γ-glutamyl Cu-binding peptide from Schizosacchromyces pombe. J. Biol Chem 263: 4186-4192 Reese RN, white CA, Winge DR (1987) Cadmium-sulfide crystallites in Cd-( -EC)nG peptide complexes form tomato. Plant Physiol 85: 1031-1035 Robinson NJ, Tommey AM, Kuske C, Jackson PJ (1993) Plant metallothioneins. Biochem J 295 ( Pt 1): 1-10 Rybakin V, Clemen CS (2005) Coronin proteins as multifunctional regulators of the cytoskeleton and membrane trafficking.. Bioessays. 2005 (6): 625-32 Rybakin V, Stumpf M, Schulze A, Majoul IV, Noegel AA, Hasse (2004) Coronin 7, the mammalian POD-1 homologue, localizes to the Golgi apparatus. A.FEBS Lett.573(1-3):161-7 S.F. Gothel, M.A. Marahiel (1999) Peptidyl-prolyl cis–trans isomerases, a superfamily of ubiquitous folding catalysts. Cell Mol. Life Sci. 55: 423–436 109 Saitoh, M., Nishitoh, H., Fujii, M., Takeda, K., Tobiume, K., Sawada, Y., Kawabata, M., Miyazono, K., Ichijo, H. (1998) Mammalian thioredoxin is a direct inhibitor of apoptosis signal-regulating kinase (ASK) 1, EMBO J. 17: 2596-2606 Sauge-Merle S, Cuine S, Carrier P, Lecomte-Pradines C, Luu DT, Peltier G (2003) Enhanced toxic metal accumulation in engineered bacterial cells expressing Arabidopsis thaliana phytochelatin synthase. Appl Environ Microbiol 69: 490-494 Shery, B. (1998)Role of cyclophilin A in the uptake of HIV-1 by macrophage and T lymphocytes.Proc. Natl. Acad. Sci. U.S.A. 95: 1758-1763 Sigal N.H, Dumont F.J (1992) Annu. Rev. Immunol. 10: 519–560 Spyrou, G., Enmark, E., Miranda-Vizuete, A., and Gustafsson, J. (1997) Cloning and Expression of a Novel Mammalian Thioredoxin, J. Biol. Chem. 272: 2936-2941 T. Nagata et al., (2000) Possible involvement of cyclophilin B and caspase-activated deoxyribonuclease in the induction of chromosomal DNA degradation in TCR-stimulated thymocytes. J. Immunol. 165: 4281–4289 Tanaka, T, Hosoi, F, Yamaguchi-Iwai, Y, Nakamura, H, Masutani, H, Udea, S, Nishiyama, A., Takeda, S, Wada, H, Spyrou, G, Yodoi, J. (2002) Thioredoxin-2 (TRX-2) is an essential gene regulating mitochondria-dependent apoptosis EMBO, J. 21: 1695-1703 Thumann J, Grill E, Winnacker E-L, Zenk MH (1991) Reactivation of metal-requiring apoenzymes by phytochelatin-metal complexes. FEBS Lett 184: 66-69 Tynecka Z, Szczes niak Z (1991) Effect of Cd2+ on phosphate uptake by cadmiun-resistance and cadmium-sensitive Staphylococcus aureus. Microbios 68: 53-63 Vogeli-Lange R, Wagner GJ (1990) Subcellular localization of cadmium and cadmium-binding peptides in tobacco leaves. Plant Physiol 92: 1086-1093 Watabe S., Hiroi T., Yamamoto Y., Fujioka Y., Hasegawa H., Yago N., Takahashi SY (1997) SP-22 is a thioredoxin-dependent peroxide reductase in mitochondria, Eur. J. Biochem. 249: 52-60 Yan Chen , Jiyang Cai§¶, T. J. Murphy , Dean P. Jones (2002) Overexpress Human Mitochondrial Thioredoxin Confers Resistance to Oxidant-induced Apoptosis in Human Osterosarcoma Cells. J. Biol. Chem., 277 (36): 33242-33248 2002 Yazykova MY, Danshina PV, Schmalhausen EV, Muronetz VI (2000) Participation of glyceraldehyde-3-phosphate dehydrogenase in the regulation of 2,3-diphosphoglycerate level in erythrocytes. Fokina KV,. Biochemistry (Mosc). 110 65 (4): 463-8 Martynowicz H, Skoczynska A, Wojakowska A Turczyn B (2004).Serum vasoactive agents in rats poisoned with cadmium. Int J Occup Med Environ Health. 17 (4):479-85 Rajalakshmi .S and Mohandas .A Copper-induced changes in tissue enzyme activity in a freshwater mussel .Ecotoxicoligy and Environmental Safety 62 (1): 140-143 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36636 | - |
| dc.description.abstract | 裂殖性酵母菌 Schizosaccharomyces pombe ATCC 2476 生長在 1 mM 鎘濃度環境下，仍能維持細胞生長。經由 HPLC 分析結果，得知 ATCC 2476 在鎘的誘導下，並不會產生 PC 與鎘螯合。將其細胞粗抽液經一連串純化步驟與製備式電泳溶離，再以二維圖譜分離，挖出可與鎘結合的若干色點進行質譜分析 LC-MS/MS，所得到蛋白質身分，與基因庫資料序列比對，得知所誘生之鎘結合蛋白質為：cyclophilin, nuclear transport factor 2 (NTF2), cyclin-dependent kinase regulatory subunit (CKS), 2,3-bisphosphoglycerate -dependent phosphoglycerate mutase, thioredoxin (Trx1) 及WD 蛋白質。推測當裂殖性酵母菌細胞受到外來鎘離子逆境時，thioredoxin 先與鎘離子結合：當過多鎘離子進入細胞質，會在細胞質中產生大量自由基，此時具抗氧化能力之 thioredoxin 及 cyclophilin 可能會形成第二道防線消滅自由基。 | zh_TW |
| dc.description.abstract | Fission yeast (Schizosaccharomyces pombe ATCC 2476) survives the toxic growing condition of 1 mM Cd. However, the HPLC analysis showed that it does not produce the heavy metal binding peptides, phytochelatins. The crude extract of the fission yeast was purified by a series of chromatographic column, and the final step of preparative electrophoresis; several proteins which could bind Cd were further isolated from 2-dimensional electrophoresis, and then subjected to LC/MS/MS. These proteins were identified as cyclophilin, nuclear transport factor 2 (NTF2), cyclin-dependent kinase regulatory subunit (CKS), 2,3-bisphosphoglycerate- dependent phosphoglycerate mutase, thioredoxin (Trx1) and WD protein. It is proposed that the fission yeast might chelate the invading Cd by its thioredoxin first, then the free radical might increase and will be removed by thioredoxin and clyclophilin if the environmental Cd keeps increasing, which serve as the second defense line. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T08:08:51Z (GMT). No. of bitstreams: 1 ntu-94-R91623608-1.pdf: 2375038 bytes, checksum: 9e14aa87a6b59aff03652476b204ab63 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 中文摘要.................................................................................................................................................4
英文摘要.................................................................................................................................................5 第一章 緒 論.........................................................................................................................................6 1.1 鎘污染..........................................................................................................................................6 1.1.1 重金屬的定義.......................................................................................................................6 1.1.2 重金屬對生物體所造成的傷害...........................................................................................7 1.1.3 鎘.........................................................................................................................................8 1.2 生物的重金屬解毒和容忍機制.................................................................................................10 1.2.1 金屬硫蛋白.........................................................................................................................10 1.2.2 植物螯合素.........................................................................................................................13 1.3 植物螯合素之合成.....................................................................................................................14 1.3.1 植物螯合素與鎘結合形成複合體.....................................................................................15 1.4 重金屬污染的解決.....................................................................................................................18 1.5 實驗緣起及研究目標.................................................................................................................20 1.5.1 酵母菌的生長觀察.............................................................................................................20 1.5.2 RP-HPLC 層析圖譜 PC 的變化...................................................................................20 1.5.3 實驗的構想與目的.............................................................................................................21 第二章 材料與方法..............................................................................................................................23 2.1 酵母菌培養................................................................................................................................23 2.1.1 菌種...................................................................................................................................23 2.1.2 洋菜膠培養法.....................................................................................................................23 2.1.3 液態培養法.........................................................................................................................25 2.2 酵母菌的粗抽取液.....................................................................................................................27 2.2.1 玻璃砂大量萃取法.............................................................................................................27 2.3 生物化學分析法.........................................................................................................................30 2.3.1 蛋白質定量法.....................................................................................................................30 2.3.2 二次元電泳之蛋白質定量方法.........................................................................................31 2.3.3 胺基酸組成分析法.............................................................................................................33 2.3.4 蛋白質N-端序列決定法....................................................................................................34 2.3.5 LC-MS/MS蛋白質序列決定法...........................................................................................36 2.3.6 鎘離子的定測.....................................................................................................................38 2.4 一般化學檢定法.........................................................................................................................40 2.4.1 硫醇基的測定.....................................................................................................................40 2.5 管柱層析法................................................................................................................................43 2.5.5 高效液相層析法 (HPLC) 基本操作................................................................................43 2.6 電泳檢定法................................................................................................................................46 2.6.1 SDS 膠體電泳......................................................................................................................46 3 2.6.2 Tricine SDS-PAGE ...............................................................................................................51 2.6.3 二維電泳............................................................................................................................57 2.6.5 膠體染色法.........................................................................................................................61 2.6.6 膠片乾燥法.........................................................................................................................65 2.7 鎘結合蛋白質的純化法.............................................................................................................67 2.7.1 膠體過濾法.........................................................................................................................67 2.7.2 離子交換法.........................................................................................................................70 2.7.3 製備式電泳與電泳溶離.....................................................................................................71 第三章 結果與討論..............................................................................................................................75 3.1 植物螯合素的檢定.....................................................................................................................75 3.2 鎘誘生蛋白質.............................................................................................................................76 3.3 鎘結合蛋白質的純化.................................................................................................................78 3. 3.1 膠體過濾層析法 (Sephadex G-50)...................................................................................78 3.3.2 離子交換層析法.................................................................................................................79 3.3.3 製備式電泳及檢測鎘結合蛋白質.....................................................................................79 3.3.4 二維電泳分析鎘結合蛋白.................................................................................................80 3.4 蛋白質定序結果分析.................................................................................................................81 3.5 定序蛋白質性質分析.................................................................................................................82 3.5.1 Cyclophilin .....................................................................................................................82 3.5.2 Nuclear transport factor 2 (NTF2) .................................................................................83 3.5.3 Cyclin-dependent kinase regulatory subunit (CKS)........................................................84 3.5.4 2,3-bisphosphoglycerate-dependent phosphoglycerate mutase......................................84 3.5.5 Thioredoxin (Trx1)..........................................................................................................85 3.5.6 WD.................................................................................................................................85 第四章 結 論.....................................................................................................................................103 4.1 結論..........................................................................................................................................103 4.2 未來展望..................................................................................................................................105 問答錄................................................................................................................................................. 110 | |
| dc.language.iso | zh-TW | |
| dc.subject | 境 | zh_TW |
| dc.subject | 鎘 | zh_TW |
| dc.subject | 逆 | zh_TW |
| dc.subject | metal | en |
| dc.subject | stress | en |
| dc.subject | Heavy | en |
| dc.title | 裂殖性酵母菌 S. pombe 2476 鎘誘導蛋白質之
純化與性質檢定 | zh_TW |
| dc.title | Purification and Characterization of
Cadmium-induced Protein from Fission Yeast Schizosaccharomyces pombe 2476 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃鵬林(Pung-Lin Huang),楊健志(Chien-Chih Yang),陳翰民(Han-Ming Chen) | |
| dc.subject.keyword | 鎘,逆,境, | zh_TW |
| dc.subject.keyword | Heavy,metal,stress, | en |
| dc.relation.page | 77 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-21 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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