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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 靳宗洛(Tsung-Luo Jinn) | |
dc.contributor.author | Ching-Chih Lin | en |
dc.contributor.author | 林敬智 | zh_TW |
dc.date.accessioned | 2021-06-17T01:48:52Z | - |
dc.date.available | 2022-08-01 | |
dc.date.copyright | 2017-08-01 | |
dc.date.issued | 2017 | |
dc.date.submitted | 2017-07-25 | |
dc.identifier.citation | Abreu I, Cabelli D (2010) Superoxide dismutases-a review of the metal-associated mechanistic variations. Biochim Biophys Acta. 1804: 263-274
Anastassia V, Wolfram M-K, Thomas M, Annette R, Bernt K, Jekaterina K, Rannar S, Peep P (2007) Oxidative switches in functioning of mammalian copper chaperone Cox17. Biochem J. 408: 139-148 Andreini C, Bertini I, Cavallaro G, Holliday G, Thornton J (2008) Metal ions in biological catalysis: from enzyme databases to general principles. J Biol Inorg Chem 13: 1205-1218 Andrés-Colás N, Perea-García A, Mayo de Andrés S, Garcia-Molina A, Dorcey E, Rodríguez-Navarro S, Pérez-Amador M, Puig S, Peñarrubia L (2013) Comparison of global responses to mild deficiency and excess copper levels in Arabidopsis seedlings. Metallomics. 5: 1234-1246 Audrey LL, Andrew ST, Thomas VOH, Amy CR (2001) Heterodimeric structure of superoxide dismutase in complex with its metallochaperone. Nature Structural Biology 8: 751-755 Banci L, Bertini I, Cantini F, Felli I, Gonnelli L, Hadjiliadis N, Pierattelli R, Rosato A, Voulgaris P (2006) The Atx1-Ccc2 complex is a metal-mediated protein-protein interaction. Nat Chem Biol. 2: 367-368 Banci L, Bertini I, Cantini F, Kozyreva T, Massagni C, Palumaa P, Rubino J, Zovo K (2012) Human superoxide dismutase 1 (hSOD1) maturation through interaction with human copper chaperone for SOD1 (hCCS). Proc Natl Acad Sci U S A. 109: 13555-13560 Banci L, Bertini I, Cramaro F, Del Conte R, Viezzoli M (2003) Solution structure of Apo Cu,Zn superoxide dismutase: role of metal ions in protein folding. Biochemistry. 42: 9543-9553 Bertini I, Hartmann H, Klein T, Liu G, Luchinat C, Weser U (2000) High resolution solution structure of the protein part of Cu7 metallothionein. Eur J Biochem. 267: 1008-1018 Blasco F, Dos-Santos J, Magalon A, Frixon C, Guigliarelli B, Santini C, Giordano G (1998) NarJ is a specific chaperone required for molybdenum cofactor assembly in nitrate reductase A of Escherichia coli. Mol Microbiol. 28: 435-447 Branislav R-N, Lukas N, Jaromir G, Ondrej Z, Michal M, Tomas E, Marie S, Vojtech A, Rene K (2013) The Role of Metallothionein in Oxidative Stress. Int J Mol Sci. 14: 6044-6066 Brose J, La Fontaine S, Wedd AG, Xiao Z (2014) Redox sulfur chemistry of the copper chaperone Atox1 is regulated by the enzyme glutaredoxin 1, the reduction potential of the glutathione couple GSSG/2GSH and the availability of Cu(I). Metallomics 6: 793-808 Brown N, Torres A, Doan P, O'Halloran T (2004) Oxygen and the copper chaperone CCS regulate posttranslational activation of Cu,Zn superoxide dismutase. Proc Natl Acad Sci U S A. 101: 5518-5523 Cai L, Klein J, Kang Y (2000) Metallothionein inhibits peroxynitrite-induced DNA and lipoprotein damage. J Biol Chem. 275: 38957-38960 Carroll. MC, Girouard. JB, Ulloa. JL, Subramaniam. JR, Wong. PC, Valentine. JS, Culotta VC (2004) Mechanisms for activating Cu- and Zn-containing superoxide dismutase in the absence of the CCS Cu chaperone. Proc Natl Acad Sci U S A. 101: 5964-5969 Chu C-C, Lee W-C, Guo W-Y, Pan S-M, Chen L-J, Li H-m, Jinn T-L (2005) A Copper Chaperone for Superoxide Dismutase That Confers Three Types of Copper/Zinc Superoxide Dismutase Activity in Arabidopsis. Plant Physiology 139: 425-436 Ciriolo M, Battistoni A, Falconi M, Filomeni G, Rotilio G (2001) Role of the electrostatic loop of Cu,Zn superoxide dismutase in the copper uptake process. Eur J Biochem. 263: 737-742 Ciriolo M, Desideri A, Paci M, Rotilio G (1990) Reconstitution of Cu,Zn-superoxide dismutase by the Cu(I).glutathione complex. J Biol Chem. 265: 11030-11034 Colpas G, Brayman T, Ming L, Hausinger R (1999) Identification of metal-binding residues in the Klebsiella aerogenes urease nickel metallochaperone, UreE. Biochemistry. 38: 4078-4088 Culotta. VC, Klomp. LWJ, Strain. J, Casareno. RLB, Krems. B, Gitlin. JD (1997) The Copper Chaperone for Superoxide Dismutase. J Biol Chem. 272: 23469-23472 Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible WR (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139: 5-17 Das A, Plotkin S (2013) SOD1 exhibits allosteric frustration to facilitate metal binding affinity. Proc Natl Acad Sci U S A. 110: 3871-3876 Ferreira AM, Ciriolo MR, Marcocci L, Rotilio G (1993) Copper(I) transfer into metallothionein mediated by glutathione. Biochem J. 292: 673-676 Gamonet F, Lauquin G (1998) The Saccharomyces cerevisiae LYS7 gene is involved in oxidative stress protection. Eur J Biochem. 251: 716-723 Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem. 48: 909-930 Graham N, Guillaume Q, Amna M, Sejir C, Christine HF (2011) Glutathione. Arabidopsis Book. 9 Gunter T, Gerstner B, Gunter K, Malecki J, Gelein R, Valentine W, Aschner M, Yule D (2013) Manganese transport via the transferrin mechanism. Neurotoxicology. 34: 118-127 Guo W-J, Meetam M, Goldsbrough PB (2008) Examining the Specific Contributions of Individual Arabidopsis Metallothioneins to Copper Distribution and Metal Tolerance. Plant Physiol. 146: 1697-1706 Huang C, Kuo W, Weiss C, Jinn T (2012) Copper chaperone-dependent and -independent activation of three copper-zinc superoxide dismutase homologs localized in different cellular compartments in Arabidopsis. Plant Physiol. 158: 737-746 Irving H, Williams RJP (1953) The stability of transition-metal complexes. J. Chem. Soc.: 3192-3210 Kevin WS, Yuewei S, Herman LL, Lindsay KB, Armando D, Valentine. JS, Edith BG (2013) Yeast copper–zinc superoxide dismutase can be activated in the absence of its copper chaperone. J Biol Inorg Chem. 18: 985-992 Khatai L, Goessler W, Lorencova H, Zangger K (2004) Modulation of nitric oxide-mediated metal release from metallothionein by the redox state of glutathione in vitro. Eur J Biochem. 27: 2408-2416 Kim D-O, Chun OK, Kim YJ, Moon H-Y, Lee CY (2003) Quantification of Polyphenolics and Their Antioxidant Capacity in Fresh Plums. J. Agric. Food Chem 51: 6509–6515 Kliebenstein D, Monde R, Last R (1998) Superoxide dismutase in Arabidopsis: an eclectic enzyme family with disparate regulation and protein localization. Plant Physiol. 118: 637-650 Krezel A, Maret W (2007) Dual nanomolar and picomolar Zn(II) binding properties of metallothionein. J Am Chem Soc. 129: 10911-10921 Krumova K, Cosa G (2016) Overview of Reactive Oxygen Species. In S Nonell, C Flors, eds, Singlet Oxygen : Applications in Biosciences and Nanosciences, Vol 1. Royal Society of Chemistry Kung C, Huang W, Huang Y, Yeh K (2006) Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry. Proteomics. 6: 2746-2758 Kuo WY, Huang CH, Liu AC, Cheng CP, Li SH, Chang WC, Weiss C, Azem A, Jinn TL (2013) CHAPERONIN 20 mediates iron superoxide dismutase (FeSOD) activity independent of its co-chaperonin role in Arabidopsis chloroplasts. New Phytol 197: 99-110 Lamb A, Torres A, O'Halloran T, Rosenzweig A (2001) Heterodimeric structure of superoxide dismutase in complex with its metallochaperone. Nat Struct Biol. 8: 751-755 Lamb A, Wernimont A, Pufahl R, Culotta V, O'Halloran T, Rosenzweig A (1999) Crystal structure of the copper chaperone for superoxide dismutase. Nat Struct Biol. 6: 724-729 Lamb AL, Torres AS, O’Halloran1 TV, Rosenzweig AC (2001) Heterodimeric structure of superoxide dismutase in complex with its metallochaperone. nature structural biology 8: 751-755 Leitch J, Jensen L, Bouldin S, Outten C, Hart P, Culotta V (2009) Activation of Cu,Zn-superoxide dismutase in the absence of oxygen and the copper chaperone CCS. J Biol Chem. 284: 21863-21871 Lepock JR, Arnold LD, Petkau A, Kelly K (1981) INTERACTION OF SUPEROXIDE DISMUTASE WITH PHOSPHOLIPID LIPOSOMES AN UPTAKE, SPIN LABEL AND CALORIMETRIC STUDY. Biochimica et Biophysica Acta 649: 45-57 Leszczyszyn O, Imam H, Blindauer C (2013) Diversity and distribution of plant metallothioneins: a review of structure, properties and functions. Metallomics. 5: 1146-1169 Lin S, Culotta V (1996) Suppression of oxidative damage by Saccharomyces cerevisiae ATX2, which encodes a manganese-trafficking protein that localizes to Golgi-like vesicles. Mol Cell Biol. 16: 6303-6312 Lindberg M, Normark J, Holmgren A, Oliveberg M (2004) Folding of human superoxide dismutase: disulfide reduction prevents dimerization and produces marginally stable monomers. Proc Natl Acad Sci 101: 15893-15898 Liu S, Fabisiak J, Tyurin V, Borisenko G, Pitt B, Lazo J, Kagan V (2000) Reconstitution of apo-superoxide dismutase by nitric oxide-induced copper transfer from metallothioneins. Chem Res Toxicol. 13: 922-931 Ma XB, Yang J (2011) An optimized preparation method to obtain high-quality RNA from dry sunflower seeds. Genet Mol Res 10: 160-168 Maret W, Vallee B (1998) Thiolate ligands in metallothionein confer redox activity on zinc clusters. Proc Natl Acad Sci U S A. 95: 3478-3482 Mark CC, Caryn EO, Jody BP, Leah R, Walter HW, Lisa JW, Hart. PJ, Laran TJ, Valeria CC (2006) The Effects of Glutaredoxin and Copper Activation Pathways on the Disulfide and Stability of Cu,Zn Superoxide Dismutase. J Biol Chem. 281: 28648-28656 Mattle D, Zhang L, Sitsel O, Pedersen L, Moncelli M, Tadini-Buoninsegni F, Gourdon P, Rees D, Nissen P, Meloni G (2015) A sulfur‐based transport pathway in Cu+‐ATPases. EMBO Rep. 16: 728-740 Miller A (2012) Superoxide dismutases: ancient enzymes and new insights. FEBS Lett. 586: 585-595 Mir G, Domènech J, Huguet G, Guo W, Goldsbrough P, Atrian S, Molinas M (2004) A plant type 2 metallothionein (MT) from cork tissue responds to oxidative stress. J Exp Bot. 55: 2488-2493 Monosson E (2012) Evolution in a Toxic World. Springer Murphy L, Strange R, Hasnain S (1997) A critical assessment of the evidence from XAFS and crystallography for the breakage of the imidazolate bridge during catalysis in CuZn superoxide dismutase. Structure. 5: 371-379 Myouga F, Hosoda C, Umezawa T, Iizumi H, Kuromori T, Motohashi R, Shono Y, Nagata N, Ikeuchi M, Shinozaki K (2008) A heterocomplex of iron superoxide dismutases defends chloroplast nucleoids against oxidative stress and is essential for chloroplast development in Arabidopsis. Plant Cell. 20: 3148-3162 Nedd S, Redler R, Proctor E, Dokholyan N, Alexandrova A (2014) Cu,Zn-superoxide dismutase without Zn is folded but catalytically inactive. J Mol Biol. 426: 4112-4124 Philpott C (2012) Coming into view: eukaryotic iron chaperones and intracellular iron delivery. J Biol Chem. 287: 13518-13523 Pufahl R, Singer C, Peariso K, Lin S, Schmidt P, Fahrni C, Culotta V, Penner-Hahn J, O'Halloran T (1997) Metal Ion Chaperone Function of the Soluble Cu(I) Receptor Atx1. Science. 278: 853-856 Pufahl RA, Singer CP, Peariso KL, Lin S-J, Schmidt PJ, Fahrni CJ, Culotta VC, Penner-Hahn JE, O’Halloran TV (1997) Metal Ion Chaperone Function of the Soluble Cu(I) Receptor Atx1. Science 278: 853-856 Puig S, Mira H, Dorcey E, Sancenón V, Andrés-Colás N, Garcia-Molina A, Burkhead J, Gogolin K, Abdel-Ghany S, Thiele D, Ecker J, Pilon M, Peñarrubia L (2007) Higher plants possess two different types of ATX1-like copper chaperones. Biochem Biophys Res Commun. 354: 385-390 R.Benatti M, Yookongkaew N, Meetam M, Guo W, Punyasuk N, AbuQamar S, Goldsbrough P (2014) Metallothionein deficiency impacts copper accumulation and redistribution in leaves and seeds of Arabidopsis. New Phytol. 202: 940-951 Rae T, Schmidt P, Pufahl R, Culotta V, O'Halloran T (1999) Undetectable intracellular free copper: the requirement of a copper chaperone for superoxide dismutase. Science. 284: 805-808 Redler R, Wilcox K, Proctor E, Fee L, Caplow M, Dokholyan N (2011) Glutathionylation at Cys-111 Induces Dissociation of Wild Type and FALS Mutant SOD1 Dimers. Biochemistry. 50: 7057-7066 Robinson N, Winge D (2010) Copper metallochaperones. Annu Rev Biochem. 79: 537-562 Rosenzweig A, Huffman D, Hou M, Wernimont A, Pufahl R, O'Halloran T (1999) Crystal structure of the Atx1 metallochaperone protein at 1.02 A resolution. Structure. 8: 605-617 Schafer F, Buettner G (2001) Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. Free Radic Biol Med. 30: 1191-1212 Schmidt P, Rae T, Pufahl R, Hamma T, Strain J, O'Halloran T, Culotta V (1999) Multiple protein domains contribute to the action of the copper chaperone for superoxide dismutase. J Biol Chem. 274: 23719-23725 Sea. K, Sohn. SH, Durazo. A, Sheng. Y, Shaw. BF, Cao. X, Taylor. AB, Whitson. LJ, Holloway. SP, Hart. PJ, Cabelli. DE, Gralla. EB, Valentine. JS (2015) Insights into the Role of the Unusual Disulfide Bond in Copper-Zinc Superoxide Dismutase. J Biol Chem. 290: 2405-2418 Shi W, Chance M (2008) Metallomics and metalloproteomics. Cell Mol Life Sci. 65: 3040-3048 Shi Y, Mowery R, Shaw B (2013) Effect of metal loading and subcellular pH on net charge of superoxide dismutase-1. J Mol Biol. 425: 4388-4404 Shin LJ, Lo JC, Yeh KC (2012) Copper chaperone antioxidant protein1 is essential for copper homeostasis. Plant Physiol 159: 1099-1110 Shin. DS, DiDonato. M, Barondeau. DP, Hura. GL, Hitomi. C, Berglund. JA, Getzoff. ED, Cary. SC, Tainer JA (2009) Superoxide Dismutase Structures, Stability, Mechanism and Insights into the Human Disease Amyotrophic Lateral Sclerosis from Eukaryotic Thermophile Alvinella pompejana. J Mol Biol. 385: 1534-1555 Silke L, Werner K (1999) Role of XDHC in Molybdenum Cofactor Insertion Into Xanthine Dehydrogenase of Rhodobacter Capsulatus. J Bacteriol. 181: 2745-4751 Smith S, She Y, Roberts E, Sarkar B (2004) Using immobilized metal affinity chromatography, two-dimensional electrophoresis and mass spectrometry to identify hepatocellular proteins with copper-binding ability. J Proteome Res. 3: 834-840 Song Y, Zhang H, Chen C, Wang G, Zhuang K, Cui J, Shen Z (2014) Proteomic analysis of copper-binding proteins in excess copper-stressed rice roots by immobilized metal affinity chromatography and two-dimensional electrophoresis. Biometals. 27: 265-276 Speisky H, Gómez M, Burgos-Bravo F, López-Alarcón C, Jullian C, Olea-Azar C, Aliaga M (2009) Generation of superoxide radicals by copper-glutathione complexes: redox-consequences associated with their interaction with reduced glutathione. Bioorg Med Chem. 17: 1803-1810 Stohs SJ, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med. 18: 321-336 Strange RW, Antonyuk S, Hough MA, Doucette PA, Rodriguez JA, Hart PJ, Hayward LJ, Valentine JS, Hasnain SS (2003) The Structure of Holo and Metal-deficient Wild-type Human Cu, Zn Superoxide Dismutase and its Relevance to Familial Amyotrophic Lateral Sclerosis. Journal of Molecular Biology 328: 877-891 Svenja P, Dana K, Michael K, Artur P, Dingeldein., Moritz S, Niemiec., Jörgen Å, Pernilla W-S (2015) Human Cytoplasmic Copper Chaperones Atox1 and CCS Exchange Copper Ions in Vitro. Biometals. 28: 577-585 Tan Y-F, Nicholas OT, Nicolas LT, Millar AH (2010) Divalent Metal Ions in Plant Mitochondria and Their Role in Interactions with Proteins and Oxidative Stress-Induced Damage to Respiratory Function. Plant Physiol. 152: 747-761 Thornalley P, Vasák M (1985) Possible role for metallothionein in protection against radiation-induced oxidative stress. Kinetics and mechanism of its reaction with superoxide and hydroxyl radicals. Biochim Biophys Acta. 827: 36-44 Williams RJP (2001) Chemical selection of elements by cells. Coordination Chemistry Reviews 216–217: 583-595 Yamakura F, Kawasaki H (2010) Post-translational modifications of superoxide dismutase. Biochim Biophys Acta. 1804: 318-325 Yang J, Zhang Y (2015) Protein Structure and Function Prediction Using I-TASSER. Curr Protoc Bioinformatics 52 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/67770 | - |
dc.description.abstract | 超氧歧化酶(Superoxide Dismutase;SOD),可將超氧分子(O2.-)轉變為過氧化氫(H2O2)及氧分子(O2),具有解除氧化逆境的功能。對於銅鋅超氧歧化酶(CuZnSOD或CSD)的活化機制,目前已知有兩條路徑,一者是藉由銅鑲嵌輔助蛋白(Copper Chaperone of SOD1;CCS)的幫助,達到銅離子鑲嵌與形成內生性雙硫鍵的活化型態; 另一者在沒有CCS情況下,CSD仍然具有少量的活性,且榖胱甘肽(Glutathione)參與在非CCS的活化CuZnSOD的路徑上,但其活化途徑與機制仍未明朗。本篇論文中,利用巰基(R-SH)阻斷劑處理,進一步證實Glutathione作為氧化還原緩衝劑,透過對於金屬鑲嵌蛋白(Metalloprotein)上巰基氧化的轉譯後修飾機制,參與在非CCS的活化路徑中。此外,亦利用硫酸銨沉澱、逆向色層分析法配合巰基差異性標定與液態層析質譜儀,篩選出金屬硫蛋白(Metallothionein)可做為CCS活化過程中的銅離子來源,並且延伸探討Glutathione催化氧化還原調控在Metallothionein和金屬伴護蛋白(Metallochaperone)的金屬結合當量化學。綜合本研究之結果,說明了當金屬誘發氧化逆境發生時,細胞內銅離子相關的抗氧化逆境機制如何複雜且精密的運作,並且對於銅離子分布提供重要的金屬傳遞機制,進一步去闡述CuZnSOD如何活化於CCS或非CCS相關的路徑。 | zh_TW |
dc.description.abstract | Superoxide dismutases (SODs, EC 1.15.1.1) are the enzymes that scavenge superoxide radicals(O2.-) to protect cells from oxidative damage. The major pathway for copper/zinc superoxide dismutase (CuZnSOD or CSD) activation involves the function of a copper chaperone for SOD (CCS). Besides, the minor CCS-independent pathway remains unclear mechanism. Glutathione had been reported cooperatively assisting for CSD activation and reconstitution but the mechanism is still unclear. In this study, we aimed to elucidate how the glutathione and the metalloproteins cooperate on the activation for enzyme, especially on Metal Incorporation into Superoxide Dismutase System (MISS). Through thiol block agent treatment, we elucidate that glutathione would involve CCS-independent activation pathway as a redox buffer via sulfur-based oxidative post-translational modification. Moreover, reverse-phase high performance liquid chromatography (HPLC) and LC-MS/MS analyses were performed to identify MT2A, MT2B as a candidate. The reconstitution assay showed MT2A and MT2B providing copper storage as a source for CCS-dependent activation. The glutathione-catalyzed redox regulation experiment fine-tunes the mechanism for glutathione-mediated oxidative switch of copper binding stoichiometry. In summary, our results elucidate the copper-dependent antioxidation process on CSD activation while copper toxicity stress occurring. The findings provide important information for how copper ion trafficking in metalloproteins and potential mechanism for CCS-dependent and CCS-independent activation. | en |
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dc.description.tableofcontents | 摘要 I
Abstract II Table of Contents III Introduction 1 Metal Ions in Biological Systems 1 Reactive Oxygen Species (ROS), Oxidative Stress and Antioxidation 2 Superoxide Dismutase (SOD): Classification, Function and Characterization 5 Studies on Copper/Zinc-superoxide Dismutase (CSD) Activation 7 Metallochaperone in Metallomics: Diversity and Characterization 10 Objectives and Current Findings 13 Materials and methods 15 Plants, Bacteria, Yeasts and Growth Conditions 15 Chemicals and Antibodies 16 Seedling and Seed RNA Preparation, cDNA Synthesis, RT-PCR and Real-Time Quantitative PCR (q-PCR) 16 Total Protein Extraction, In-gel/Liquid SOD Activity Assay and Immunoblotting 17 Coomassie Blue and Silver Staining Method 18 Construct generation 19 Lysine-independent Aerobic Growth Assay 21 Recombinant Proteins Purification and Deactivation 22 Size Exclusion Chromatography for Protein Oligomer Statement Analysis 24 Reconstitution of apo-CSD1 and in vitro Treatments 24 Copper-immobilized Metal Affinity Chromatography and Gel-filtration Chromatography 27 Ammonium Sulfate (AS) Fractionation and Reverse-phase HPLC 28 Differential Labelling, Trypsin Digestion and Protein Identification 29 In vitro Pull Down Assay 31 Root Growth and Elongation Assay 31 MDA and Anthocyanin Content Quantification 32 Statistical Analysis 33 Accession Numbers 33 Results 35 Glutathione Is Involving in Arabidopsis apo-CSD1 Reconstitution as Forming the Cuprous Complex. 35 Glutathione Involved in Arabidopsis apo-CSD1 Reconstitution Reacting with the Thiol Group. 36 Glutathione Reacts with Cellular Components in apo-CSD1 Reconstitution via Thiol Group. 36 Excessive Glutathione Would Inhibit CCS-Independent Activation Pathway 37 Redox Sulfur Regulation Involved in Ion Binding Capacity via Active Cysteinyl Thiol Group 38 The Effect of Ascorbic Acid on apo-CSD1 reconstitution 39 The Metalloprotein Candidates Reconstitute apo-CSD via Non-specific Activation Process 40 The Cu(II) May not the Copper Donor Form for apo-CSD1 Reconstitution 41 Small Molecular Cysteine-rich Protein Would be Copper Source for apo-CSD1 Reconstitution 42 Apo-CSD1 Reconstitution via Metallochaperone Is Contributed by Glutathione-Catalyzed Redox Regulation on Metalloprotein 44 CCS ATX1-like Domain Is Critical for Copper Trafficking 46 CCS Interact with Metallochaperone to Ion Exchange for CSD Activation/De-activation 48 CSD Is Primary Enzymatic Antioxidant under Copper Stress and Its Activation Is Corresponding to Copper Content 49 Discussion 52 Cu(I) instead of Cu(II) Prefers to Be More Susceptible to Metal Ion Transmission in Biological System 52 Excessive GSSG may Cause Glutathionylation and Affect CSD Activation 55 Glutathione Would Work as Redox Buffer to Regulate Protein Function 56 Metallothioneins Could Serve as Copper Ion Pool for CSD Activation and ATX1, and CCH Could Incorporate Copper into CSD via Non-specific Process 56 Redox Posttranslational Modification on Cysteine Would Affect the Turnover of Copper Ion Trafficking in Proteinaceous Pool 58 Iron Trafficking between Metalloprotein and metallochaperone Regulate CSD Activation /De-activation Process in Response to Copper Content 60 CSD might Have Two Distinct Mechanisms for Copper Acquisition 61 Conclusions 62 Tables 63 Table 1. Result of LC-MS/MS analysis of HPLC fraction collected at 31 to 36 minutes. 63 Table 2. Result of LC-MS/MS analysis of HPLC fraction collected at 37 to 42 minutes. 64 Figures 65 Figure 1. Effect of different copper-glutathione or copper-phytochelatin complex on CSD1 reconstitution in vitro. 65 Figure 2. The combination effect of glutathione and the ccs-cellular extract on apo-CSD1 reconstitution in vitro. 66 Figure 3. The antagonistic effect of excessive glutathione on the ccs-cellular extract-mediated apo-CSD1 reconstitution in vitro. 68 Figure 4. Characterization of the component involved in ccs-cellular extract-mediated apo-CSD1 reconstitution in vitro. 69 Figure 5. Characterization of the redox regulation involved in ccs-cellular extract-mediated apo-CSD1 reconstitution in vitro. 70 Figure 6. The additive effect of ascorbic acid on the ccs-cellular extract-mediated on apo-CSD1 reconstitution in vitro. 71 Figure 7. Apo-CSD1 activation in the presence of ammonium sulfate (AS) precipitated fractions. 72 Figure 8. Apo-CSD1 activation in the presence of reverse-phase HPLC chromatography separated fractions. 73 Figure 9. The differential labelling on cysteinyl thiol group for LC-MS/MS identification. 74 Figure 10. The metalloprotein (MT2A and MT2B) and metallochaperone (ATX1 and CCH) inhibit CSD1 activity. 75 Figure 11. In vitro pull-down assay of CSD1 and metalloprotein or metallochaperone interaction. 76 Figure 12. The effect of glutathione-catalyzed redox state of metalloprotein/ metallochaperone -mediated metal ion incorporation for apo-CSD1 reconstitution in vitro. 77 Figure 13. Hypothetical model of glutathione-mediated redox regulation on metallochaperone and metalloprotein for copper incorporation into CSD1. 79 Figure 14. The effect of different domain of CCS protein and domain 1-like metallochaperone on apo-CSD1 reconstitution. 80 Figure 15. In vitro pull down assay of CSD1 and CCS different domain containing proteins interaction. 81 Figure 16. The copper ion exchange between metalloprotein/non-specific metallochaperone and specific metallochaperone CCS determine the CSD activation and de-activation. 82 Figure 17. In vitro pull down assay of CCS and the metallochaperone/metalloprotein interaction. 83 Figure 18. The antioxidant abilities of Wild-type (Col), ccs, csd1 plants under copper mild-deficient/excessive condition 84 Figure 19. The antioxidant abilities of Col-0, ccs, csd1 under copper mild-deficient to toxicity stress. 85 Figure 20. The activity of CSD and apo-CSD1 reconstitution under different copper concentration treatment in ccs-cellular extract. 86 Figure 21. The proposed model for CSD1 activation through CSD1/CCS/(MC/MP) complex. 87 Figure 22. Proposed reaction mechanism for copper activation pathway of CSD. 88 Figure 23. The summary of antioxidation process under metal-catalyzed oxidative stress. 89 Supplemental Data 90 Supplemental Figure S1. Structure elucidation of Cu/Zn-SOD 90 Supplemental Figure S2. Characterization of the T-DNA insertion knockout plant, ccs and csd1. 91 Supplemental Figure S3. Recombinant CSD1 purification. 92 Supplemental Figure S4. The re-activation and dimerization of reconstituted holo-CSD1. 93 Supplemental Figure S5. Effect of pH and antioxidant on Liquid SOD assay 94 Supplemental Figure S6. Metalloprotein mining by Yeast Lysine-Independent Aerobic Growth assay 95 Supplemental Figure S7. The workflow of post-ribosomal supernatant (PRS) preparation and copper-immobilized metal affinity chromatography(Cu-IMAC) and gel -filtration chromatography for metalloprotein mining for apo-CSD1 reconstitution in ccs cellular extract 96 Supplemental Figure S8. Apo-CSD1 activation in the presence of fractions by Cu-IMAC chromatography. 97 Supplemental Figure S9. Apo-CSD1 activation in the presence of fractions by gel -filtration chromatography. 98 Supplemental Figure S10. Recombinant GST tagged ATX1, CCH; 6xHis tagged MT2A, MT2B and non-tagged ATX1, CCH purification. 99 Supplemental Figure S11. The superoxide radical scavenging ability of metalloprotein (MT2A and MT2B) and metallochaperone (ATX1 and CCH). 100 Supplemental Figure S12. Recombinant MBP and MBP tagged ΔTP-CCS, CSD1; GST and GST tagged ΔTP-CCS, CCSD1, CCSD1D2, CCSD2, CCSD2D3, MT2A, MT2B and 6His tagged ΔTP-CCS purification 101 Supplemental Figure S13. yCCS and ySOD1 co-crystal structure. 102 Supplemental Table S1. Primer used in this study 103 References 104 | |
dc.language.iso | en | |
dc.title | 穀胱甘肽與金屬蛋白活化銅離子鑲嵌至阿拉伯芥銅鋅超氧岐化酶之研究 | zh_TW |
dc.title | Study of Glutathione and Metalloprotein Cooperation in Copper Ion Incorporation into Arabidopsis Copper/Zinc-Superoxide Dismutase | en |
dc.type | Thesis | |
dc.date.schoolyear | 105-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張英?(Ing-Feng Chang),王雅筠(Ya-Yun Wang),謝明勳(Ming-Hsiun Hsieh),葉國楨(Kuo-Chen Yeh) | |
dc.subject.keyword | 榖胱甘?,銅鋅超氧歧化?,金屬鑲嵌蛋白, | zh_TW |
dc.subject.keyword | glutathione,copper/zinc superoxide dismutase,metalloprotein., | en |
dc.relation.page | 111 | |
dc.identifier.doi | 10.6342/NTU201701982 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2017-07-26 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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