請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17223
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 常怡雍(Yee-yung Charng) | |
dc.contributor.author | Zih-teng Chen | en |
dc.contributor.author | 陳子騰 | zh_TW |
dc.date.accessioned | 2021-06-08T00:01:45Z | - |
dc.date.copyright | 2013-08-27 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-15 | |
dc.identifier.citation | Ang D, Chandrasekhar GN, Zylicz M, Georgopoulos C (1986) Escherichia coli grpE gene codes for heat shock protein B25.3, essential for both lambda DNA replication at all temperatures and host growth at high temperature. J Bacteriol 167: 25-29
Ang D, Georgopoulos C (1989) The heat-shock-regulated grpE gene of Escherichia coli is required for bacterial growth at all temperatures but is dispensable in certain mutant backgrounds. J Bacteriol 171: 2748-2755 Barthel S, Rupprecht E, Schneider D (2011) Thermostability of two cyanobacterial GrpE thermosensors. Plant Cell Physiol 52: 1776-1785 Bolliger L, Deloche O, Glick BS, Georgopoulos C, Jeno P, Kronidou N, Horst M, Morishima N, Schatz G (1994) A mitochondrial homolog of bacterial GrpE interacts with mitochondrial hsp70 and is essential for viability. EMBO J 13: 1998-2006 Borges JC, Fischer H, Craievich AF, Hansen LD, Ramos CH (2003) Free human mitochondrial GrpE is a symmetric dimer in solution. J Biol Chem 278: 35337-35344 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254 Brehmer D, Rudiger S, Gassler CS, Klostermeier D, Packschies L, Reinstein J, Mayer MP, Bukau B (2001) Tuning of chaperone activity of Hsp70 proteins by modulation of nucleotide exchange. Nat Struct Biol 8: 427-432 Bukau B, Deuerling E, Pfund C, Craig EA (2000) Getting newly synthesized proteins into shape. Cell 101: 119-122 Bukau B, Weissman J, Horwich A (2006) Molecular chaperones and protein quality control. Cell 125: 443-451 Compton LA, Johnson WC, Jr. (1986) Analysis of protein circular dichroism spectra for secondary structure using a simple matrix multiplication. Anal Biochem 155: 155-167 Garnier J, Osguthorpe DJ, Robson B (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120: 97-120 Gelinas AD, Langsetmo K, Toth J, Bethoney KA, Stafford WF, Harrison CJ (2002) A structure-based interpretation of E.coli GrpE thermodynamic properties. J Mol Biol 323: 131-142 Genevaux P, Georgopoulos C, Kelley WL (2007) The Hsp70 chaperone machines of Escherichia coli: a paradigm for the repartition of chaperone functions. Mol Microbiol 66: 840-857 Grimshaw JP, Jelesarov I, Schonfeld HJ, Christen P (2001) Reversible thermal transition in GrpE, the nucleotide exchange factor of the DnaK heat-shock system. J Biol Chem 276: 6098-6104 Grimshaw JP, Jelesarov I, Siegenthaler RK, Christen P (2003) Thermosensor action of GrpE. The DnaK chaperone system at heat shock temperatures. J Biol Chem 278: 19048-19053 Grimshaw JP, Siegenthaler RK, Zuger S, Schonfeld HJ, Z'Graggen B R, Christen P (2005) The heat-sensitive Escherichia coli grpE280 phenotype: impaired interaction of GrpE(G122D) with DnaK. J Mol Biol 353: 888-896 Groemping Y, Klostermeier D, Herrmann C, Veit T, Seidel R, Reinstein J (2001) Regulation of ATPase and chaperone cycle of DnaK from Thermus thermophilus by the nucleotide exchange factor GrpE. J Mol Biol 305: 1173-1183 Groemping Y, Reinstein J (2001) Folding properties of the nucleotide exchange factor GrpE from Thermus thermophilus: GrpE is a thermosensor that mediates heat shock response. J Mol Biol 314: 167-178 Hall AE (2010) Crop Responses to Environment. Taylor & Francis Harrison C (2003) GrpE, a nucleotide exchange factor for DnaK. Cell Stress Chaperones 8: 218-224 Harrison CJ, Hayer-Hartl M, Di Liberto M, Hartl F, Kuriyan J (1997) Crystal structure of the nucleotide exchange factor GrpE bound to the ATPase domain of the molecular chaperone DnaK. Science 276: 431-435 Hartl FU, Hayer-Hartl M (2002) Molecular chaperones in the cytosol: from nascent chain to folded protein. Science 295: 1852-1858 Hu C, Lin SY, Chi WT, Charng YY (2012) Recent gene duplication and subfunctionalization produced a mitochondrial GrpE, the nucleotide exchange factor of the Hsp70 complex, specialized in thermotolerance to chronic heat stress in Arabidopsis. Plant Physiol 158: 747-758 Huang B, Xu C (2008) Identification and characterization of proteins associated with plant tolerance to heat stress. J Integr Plant Biol 50: 1230-1237 Iba K (2002) Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance. Annu Rev Plant Biol 53: 225-245 Ikeda E, Yoshida S, Mitsuzawa H, Uno I, Toh-e A (1994) YGE1 is a yeast homologue of Escherichia coli grpE and is required for maintenance of mitochondrial functions. FEBS Lett 339: 265-268 Kelly SM, Jess TJ, Price NC (2005) How to study proteins by circular dichroism. Biochim Biophys Acta 1751: 119-139 Kubo Y, Tsunehiro T, Nishikawa S, Nakai M, Ikeda E, Toh-e A, Morishima N, Shibata T, Endo T (1999) Two distinct mechanisms operate in the reactivation of heat-denatured proteins by the mitochondrial Hsp70/Mdj1p/Yge1p chaperone system. J Mol Biol 286: 447-464 Laloraya S, Dekker PJ, Voos W, Craig EA, Pfanner N (1995) Mitochondrial GrpE modulates the function of matrix Hsp70 in translocation and maturation of preproteins. Mol Cell Biol 15: 7098-7105 Laloraya S, Gambill BD, Craig EA (1994) A role for a eukaryotic GrpE-related protein, Mge1p, in protein translocation. Proc Natl Acad Sci 91: 6481-6485 Liberek K, Marszalek J, Ang D, Georgopoulos C, Zylicz M (1991a) Escherichia coli DnaJ and GrpE heat shock proteins jointly stimulate ATPase activity of DnaK. Proc Natl Acad Sci U S A 88: 2874-2878 Liberek K, Skowyra D, Zylicz M, Johnson C, Georgopoulos C (1991b) The Escherichia coli DnaK chaperone, the 70-kDa heat shock protein eukaryotic equivalent, changes conformation upon ATP hydrolysis, thus triggering its dissociation from a bound target protein. J Biol Chem 266: 14491-14496 Lin BL, Wang JS, Liu HC, Chen RW, Meyer Y, Barakat A, Delseny M (2001) Genomic analysis of the Hsp70 superfamily in Arabidopsis thaliana. Cell Stress Chaperones 6: 201-208 Lin MG, Chen BE, Liang WC, Chou WM, Chen JH, Kuo LY, Lin LL (2010) Site-saturation mutagenesis of leucine 134 of Bacillus licheniformis nucleotide exchange factor GrpE reveals the importance of this residue to the co-chaperone activity. Protein J 29: 365-372 Liu Y-G, Huang N (1998) Efficient Amplification of Insert End Sequences from Bacterial Artificial Chromosome Clones by Thermal Asymmetric Interlaced PCR. Plant Mol Biol Rep 16: 175-175 Manavalan P, Johnson WC, Jr. (1987) Variable selection method improves the prediction of protein secondary structure from circular dichroism spectra. Anal Biochem 167: 76-85 Marshall JS, DeRocher AE, Keegstra K, Vierling E (1990) Identification of heat shock protein hsp70 homologues in chloroplasts. Proc Natl Acad Sci U S A 87: 374-378 Mayer MP, Bukau B (2005) Hsp70 chaperones: cellular functions and molecular mechanism. Cell Mol Life Sci 62: 670-684 Mogk A, Tomoyasu T, Goloubinoff P, Rudiger S, Roder D, Langen H, Bukau B (1999) Identification of thermolabile Escherichia coli proteins: prevention and reversion of aggregation by DnaK and ClpB. EMBO J 18: 6934-6949 Moro F, Muga A (2006) Thermal adaptation of the yeast mitochondrial Hsp70 system is regulated by the reversible unfolding of its nucleotide exchange factor. J Mol Biol 358: 1367-1377 Moro F, Taneva SG, Velazquez-Campoy A, Muga A (2007) GrpE N-terminal domain contributes to the interaction with Dnak and modulates the dynamics of the chaperone substrate binding domain. J Mol Biol 374: 1054-1064 Morrison SL (2001) Transformation of E. coli by electroporation. Curr Protoc Immunol Appendix 3: Appendix 3N Nakamura A, Takumi K, Miki K (2010) Crystal structure of a thermophilic GrpE protein: insight into thermosensing function for the DnaK chaperone system. J Mol Biol 396: 1000-1011 Neupert W, Brunner M (2002) The protein import motor of mitochondria. Nat Rev Mol Cell Biol 3: 555-565 Oliveira CL, Borges JC, Torriani IL, Ramos CH (2006) Low resolution structure and stability studies of human GrpE#2, a mitochondrial nucleotide exchange factor. Arch Biochem Biophys 449: 77-86 Packschies L, Theyssen H, Buchberger A, Bukau B, Goody RS, Reinstein J (1997) GrpE accelerates nucleotide exchange of the molecular chaperone DnaK with an associative displacement mechanism. Biochemistry 36: 3417-3422 Padidam M, Reddy VS, Beachy RN, Fauquet CM (1999) Molecular characterization of a plant mitochondrial chaperone GrpE. Plant Mol Biol 39: 871-881 Pain RH (2005) Determining the fluorescence spectrum of a protein. Curr Protoc Protein Sci Chapter 7: Unit 7.7 Palleros DR, Reid KL, Shi L, Welch WJ, Fink AL (1993) ATP-induced protein-Hsp70 complex dissociation requires K+ but not ATP hydrolysis. Nature 365: 664-666 Savchenko GE, Klyuchareva EA, Abramchik LM, Serdyuchenko EV (2002) Effect of Periodic Heat Shock on the Inner Membrane System of Etioplasts. Russian Journal of Plant Physiology 49: 349-359 Schmid D, Baici A, Gehring H, Christen P (1994) Kinetics of molecular chaperone action. Science 263: 971-973 Schmidt TG, Skerra A (2007) The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. Nat Protoc 2: 1528-1535 Schroda M, Vallon O, Whitelegge JP, Beck CF, Wollman FA (2001) The chloroplastic GrpE homolog of Chlamydomonas: two isoforms generated by differential splicing. Plant Cell 13: 2823-2839 Sreerama N, Woody RW (2000) Estimation of protein secondary structure from circular dichroism spectra: comparison of CONTIN, SELCON, and CDSSTR methods with an expanded reference set. Anal Biochem 287: 252-260 Su PH, Li HM (2010) Stromal Hsp70 is important for protein translocation into pea and Arabidopsis chloroplasts. Plant Cell 22: 1516-1531 Sung DY, Vierling E, Guy CL (2001) Comprehensive expression profile analysis of the Arabidopsis Hsp70 gene family. Plant Physiol 126: 789-800 Terpe K (2003) Overview of tag protein fusions: from molecular and biochemical fundamentals to commercial systems. Appl Microbiol Biotechnol 60: 523-533 Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76: 4350-4354 Voos W, Gambill BD, Laloraya S, Ang D, Craig EA, Pfanner N (1994) Mitochondrial GrpE is present in a complex with hsp70 and preproteins in transit across membranes. Mol. Cell. Biol. 14: 6627-6634 Wahid A, Gelani S, Ashraf M, Foolad MR (2007) Heat tolerance in plants: An overview. Environmental and Experimental Botany 61: 199-223 Westermann B, Pripbuus C, Neupert W, Schwarz E (1995) The Role of the Grpe Homolog, Mge1p, in Mediating Protein Import and Protein-Folding in Mitochondria. EMBO J 14: 3452-3460 Whitmore L, Wallace BA (2008) Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases. Biopolymers 89: 392-400 Wu CC, Naveen V, Chien CH, Chang YW, Hsiao CD (2012) Crystal structure of DnaK protein complexed with nucleotide exchange factor GrpE in DnaK chaperone system: insight into intermolecular communication. J Biol Chem 287: 21461-21470 Yeh CH, Kaplinsky NJ, Hu C, Charng YY (2012) Some like it hot, some like it warm: Phenotyping to explore thermotolerance diversity. Plant Sci 195: 10-23 Young JC, Agashe VR, Siegers K, Hartl FU (2004) Pathways of chaperone-mediated protein folding in the cytosol. Nat Rev Mol Cell Biol 5: 781-791 Zeng G (1998) Sticky-end PCR: new method for subcloning. Biotechniques 25: 206-208 Zhang J (2003) Evolution by gene duplication: an update. Trends in ecology & evolution 18: 292-298 Zhu X, Zhao X, Burkholder WF, Gragerov A, Ogata CM, Gottesman ME, Hendrickson WA (1996) Structural analysis of substrate binding by the molecular chaperone DnaK. Science 272: 1606-1614 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17223 | - |
dc.description.abstract | Prokaryotic GrpE acts as a nucleotide-exchange factor in the Hsp70/DnaK chaperone machinery. Two GrpE homologs were identified in Arabidopsis mitochondria (Mge1 and Mge2). Mge2, but not Mge1, restores the growth of heat-sensitive E. coli grpE mutant, DA16, at 43 oC. Although Mge1 and Mge2 are highly similar in primary structure, Mge2 possesses an extra peptide derived from a retained in-frame intron sequence. Phenotyping of Mge2 T-DNA knockout lines reveals that Mge2 is specifically required for tolerating prolonged exposure to moderately high temperature. To characterize how Mge2 confers tolerance to high temperature, plasmids expressing recombinant Mge proteins were constructed and used for complementary assay. Recombinant Mge proteins were purified and analyzed by circular dichroism. The results suggested Mge2 was more thermostable than Mge1. And the in-frame intron-derived peptide sequence of Mge2 might not affect the thermostability in vitro. Furthermore, analysis of domain-swapped Mge proteins indicated that both the long α-helix domain and the β-sheet domain might affect the thermostability of Mge proteins, but the long α-helix domain might be predominant in affecting the thermostability. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:01:45Z (GMT). No. of bitstreams: 1 ntu-102-R00b22024-1.pdf: 2756517 bytes, checksum: 56cc4d2eebcc08ea32010262cec84ced (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | Contents i
Abbreviation iv Abstract v 中文摘要 vi Figure contents vii Table contents viii Chapter I. Introduction 1 1.1 Heat stress 1 1.2 Heat shock proteins and Hsp70s 1 1.3 Mechanism of KJE chaperone cycle 2 1.4 Structure and thermodynamic properties of GrpE protein 4 1.5 GrpE homologs in eukaryotes 9 1.6 Arabidopsis Mge1 and Mge2 might be specialized for different stress 10 1.7 Objective of this study 12 Chapter II. Materials and Methods 14 2.1 Cloning of Arabidopsis Mge genes 14 2.1.1 Plasmids construction for production of Strep-tagged recombinant Mge1 and Mge2 proteins 14 2.1.2 Plasmid construction for production of recombinant Strep-tagged Mge2Δ protein 15 2.1.3 Plasmid construction for production of Strep-tagged domain-swapped Mge proteins 15 2.2 Complementary assay 18 2.3 Purification of Mge proteins 19 2.3.1 Protein expression 19 2.3.2 Cell lysis 20 2.3.3 Affinity chromatography 20 2.3.4 Ion-Exchange chromatography 21 2.3.5 Gel-filtration chromatography 22 2.4 Circular dichorism (CD) 22 2.5 Mass analysis 23 2.5.1 In-gel digestion 23 2.5.2 Zip-Tip purification 24 2.5.3 MS analysis 24 Chapter III. Results 26 3.1 The Strep-tag does not alter the function of recombinant Mge proteins. 26 3.2 Purification of recombinant Strep-tagged Mge proteins. 28 3.3 CD analysis showed that Mge2 was more thermostable than Mge1 at elevated temperatures. 29 3.4 Deletion of the intron-derived peptide sequence in Mge2 did not affect the function of Mge2 in complementation of E. coli DA16 mutant. 31 3.5 Deletion of the intron-derived peptide sequence in Mge2 did not affect protein thermostability in vitro. 32 3.6 The long α-helix domain in Mge determines the capacity in conferring tolerance to high temperature in vivo. 33 3.7 CD analysis showed that the long α-helix is the most important domain affecting Mge thermostability in vitro. 35 Chapter IV. Discussion 37 4.1 Mge1 and Mge2 might possess different affinity to DnaK. 37 4.2 The Arabidopsis Mge proteins might function similarly to S. cerevisiae Mge1p rather than E. coli GrpE. 38 4.3 Mge2 might remain active during chronic heat stress. 40 4.4 The long α-helix domain might be predominant to the thermostability of Arabidopsis Mge proteins. 41 Chapter V. Future work 44 5.1 complementation analysis of Mge2Δ in mge2 mutant plants. 44 5.2 Identification of the relationship between the thermal unfolding transition and monomerization of Mge proteins 44 5.3 Measurements of NEF activity of Mge proteins 45 References 46 Figures 52 Tables 68 Appendix 73 碩士論文口試問答摘要 83 | |
dc.language.iso | en | |
dc.title | 阿拉伯芥粒線體GrpE蛋白質之熱穩定性研究 | zh_TW |
dc.title | Thermostability Analysis of Two Arabidopsis Mitochondrial GrpE Proteins | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 楊健志(Chien-chih Yang) | |
dc.contributor.oralexamcommittee | 王愛玉,陳佩燁,黃楓婷 | |
dc.subject.keyword | 阿拉伯芥,GrpE,Mge,熱穩定性, | zh_TW |
dc.subject.keyword | Arabidopsis,GrpE,Mge,thermostability, | en |
dc.relation.page | 87 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2013-08-15 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 2.69 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。