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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李秀敏(Hsou-min Li) | |
| dc.contributor.author | Ting-Fang Chen | en |
| dc.contributor.author | 陳婷芳 | zh_TW |
| dc.date.accessioned | 2022-11-24T03:04:17Z | - |
| dc.date.available | 2021-07-08 | |
| dc.date.available | 2022-11-24T03:04:17Z | - |
| dc.date.copyright | 2021-07-08 | |
| dc.date.issued | 2021 | |
| dc.date.submitted | 2021-06-24 | |
| dc.identifier.citation | Agne, B., Infanger, S., Wang, F., Hofstetter, V., Rahim, G., Martin, M., Lee, D.W., Hwang, I., Schnell, D., and Kessler, F. (2009). A toc159 import receptor mutant, defective in hydrolysis of GTP, supports preprotein import into chloroplasts. Journal of Biological Chemistry 284, 8670-8679. Andrès, C., Agne, B., and Kessler, F. (2010). The TOC complex: preprotein gateway to the chloroplast. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research 1803, 715-723. Benz, J.P., Soll, J., and Bölter, B. (2009). Protein transport in organelles: The composition, function and regulation of the Tic complex in chloroplast protein import. The FEBS Journal 276, 1166-1176. Blankenship, R.E. (2014). Molecular mechanisms of photosynthesis. Oxford, England: Blackwell Science. Bölter, B., May, T., and Soll, J. (1998). A protein import receptor in pea chloroplasts, Toc86, is only a proteolytic fragment of a larger polypeptide. FEBS Letters 441, 59-62. Chan, K.X., Phua, S.Y., Crisp, P., McQuinn, R., and Pogson, B.J. (2016). Learning the languages of the chloroplast: retrograde signaling and beyond. Annual Review of Plant Biology 67, 25-53. Chen, Y.J., and Liao, H.K. (2003). Mass spectrometry-based proteomics. Chemistry 61, 235-247. Chu, C.C., and Li, H.m. (2011). Determining the location of an Arabidopsis chloroplast protein using in vitro import followed by fractionation and alkaline extraction. Methods in Molecular Biology 774, 339-350. Chu, C.C., Swamy, K., and Li, H.m. (2020). Tissue-specific regulation of plastid protein import via transit-peptide motifs. The Plant Cell 32, 1204-1217. Cline, K., Werner-Washburne, M., Andrews, J., and Keegstra, K. (1984). Thermolysin is a suitable protease for probing the surface of intact pea chloroplasts. Plant Physiology 75, 675-678. Dahlin, C., and Cline, K. (1991). Developmental regulation of the plastid protein import apparatus. The Plant Cell 3, 1131-1140. Hiltbrunner, A., Grünig, K., Alvarez-Huerta, M., Infanger, S., Bauer, J., and Kessler, F. (2004). AtToc90, a new GTP-binding component of the Arabidopsis chloroplast protein import machinery. Plant Molecular Biology 54, 427-440. Hirsch, S., Muckel, E., Heemeyer, F., von Heijne, G., and Soll, J. (1994). A receptor component of the chloroplast protein translocation machinery. Science 266, 1989-1992. Hölzl, G., and Dörmann, P. (2019). Chloroplast lipids and their biosynthesis. Annual Review of Plant Biology 70, 51-81. Inoue, H., Rounds, C., and Schnell, D.J. (2010). The molecular basis for distinct pathways for protein import into Arabidopsis chloroplasts. The Plant Cell 22, 1947-1960. Ivanova, Y., Smith, M.D., Chen, K., and Schnell, D.J. (2004). Members of the Toc159 import receptor family represent distinct pathways for protein targeting to plastids. Molecular Biology of the Cell 15, 3379-3392. Jarvis, P., Chen, L.J., Li, H.m., Peto, C.A., Fankhauser, C., and Chory, J. (1998). An Arabidopsis mutant defective in the plastid general protein import apparatus. Science 282, 100-103. Kubis, S., Patel, R., Combe, J., Bédard, J., Kovacheva, S., Lilley, K., Biehl, A., Leister, D., Ríos, G., and Koncz, C. (2004). Functional specialization amongst the Arabidopsis Toc159 family of chloroplast protein import receptors. The Plant Cell 16, 2059-2077. Li, H.m., and Chiu, C.C. (2010). Protein transport into chloroplasts. Annual Review of Plant Biology 61, 157-180. Perry, S.E., and Keegstra, K. (1994). Envelope membrane proteins that interact with chloroplastic precursor proteins. The Plant Cell 6, 93-105. Raines, C.A. (2003). The Calvin cycle revisited. Photosynthesis Research 75, 1-10. Richardson, L.G., and Schnell, D.J. (2020). Origins, function, and regulation of the TOC–TIC general protein import machinery of plastids. Journal of Experimental Botany 71, 1226-1238. Teng, Y.S., Chan, P.T., and Li, H.m. (2012). Differential age-dependent import regulation by signal peptides. PLoS Biology 10, e1001416. Yu, T.S., and Li, H.m. (2001). Chloroplast protein translocon components atToc159 and atToc33 are not essential for chloroplast biogenesis in guard cells and root cells. Plant Physiology 127, 90-96. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/80312 | - |
| dc.description.abstract | 我們實驗室先前的研究發現葉綠體前驅蛋白可分為三種年齡選擇性組別：第一組前驅蛋白傾向運輸至年輕的葉綠體，其中包括核酮糖小亞基前驅蛋白 (prRBCS)；第二組前驅蛋白對於年齡無偏好性；第三組前驅蛋白則傾向運輸至成熟的葉綠體，其中包括葉綠體內膜機組Tic40前驅蛋白(prTic40)和葉綠體熱休克同源前驅蛋白70-1 (prcpHsc70-1)。我們並進一步發現在導向序列上的雙陽性模體（兩個帶正電荷殘基）是調控前驅蛋白優先選擇運輸至成熟葉綠體的重要關鍵，但對於雙陽性模體的受體仍不清楚。為了找出此受體與了解年齡調控前驅蛋白運輸的分子機制，我們利用接上FLAG標籤的prTic40 (prAtTic40-FLAG)作為材料，將其導向序列上雙陽性模體兩側的殘基突變成半胱氨酸，當prAtTic40-FLAG與成熟葉綠體進行結合時，利用BMH交聯劑將接近雙陽性模體附近的半胱氨酸進行交聯，再將prAtTic40-FLAG以免疫沈澱法分離出來，此時被交聯的蛋白可通過串聯式質譜儀識別。根據我的實驗結果，受體最有可能是葉綠體外膜機組蛋白159 （Toc159，其由A、G、M三個結構域組成，以下稱為 Toc159AGM）的A結構域截斷體，也就是Toc159GM。先前許多研究中皆表明A結構域經常受到降解，但原因尚不明瞭，而我在豌豆(Pisum sativum)的葉綠體觀察到A結構域受到剪切的程度會隨著年齡而增加。另外發現prcpHsc70-1及prTic40運輸進入僅表達Toc159GM的葉綠體效率較好，而他們雙陽性模體破壞的變體prcpHsc70-1(KR2324AA)及prTic40(RK2930EE)則是運輸進入含有Toc159AGM的野生型葉綠體效率較佳。此外在體外沉降實驗中亦證實prTic40導向序列會偏好與Toc159GM結合，而prRBCS導向序列則偏好與Toc159AGM結合。綜合以上結果發現Toc159的A結構域剪切可以調控年齡對葉綠體蛋白運輸的調控。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2022-11-24T03:04:17Z (GMT). No. of bitstreams: 1 U0001-2406202112540400.pdf: 4840546 bytes, checksum: 568b1affcf87d39692989cf1f6c4a88d (MD5) Previous issue date: 2021 | en |
| dc.description.tableofcontents | 致謝 I 中文摘要 II Abstract III Contents V Introduction 1 Material and Methods 5 Plant growth conditions 5 Expression and purification of recombinant AtTic40 mutant proteins 5 Chloroplast isolation 6 Protein binding assays with BMH crosslinking 6 Gel electrophoresis and immunoblotting 7 Immunoprecipitation 7 Tandem mass spectrometry and protein identification 8 In vitro transcription and translation 8 Protein import assays 9 PCR-based mutagenesis and construction of plasmids 9 In vitro pull-down assays 10 Root length measurement 10 Results 11 Discussion 20 References 23 Figures 25 Tables 38 Appendix 40 | |
| dc.language.iso | en | |
| dc.subject | 葉綠體外膜機組蛋白159（Toc159） | zh_TW |
| dc.subject | 葉綠體 | zh_TW |
| dc.subject | 蛋白運輸 | zh_TW |
| dc.subject | 雙陽性模體 | zh_TW |
| dc.subject | Translocon at the outer envelope membrane of chloroplast 159 (Toc159) | en |
| dc.subject | Chloroplasts | en |
| dc.subject | Protein import | en |
| dc.subject | Twin-positive motif | en |
| dc.title | 以受體特定剪切達到年齡對葉綠體蛋白運輸的調控 | zh_TW |
| dc.title | Specific receptor cleavage mediates age-dependent import regulation | en |
| dc.date.schoolyear | 109-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蔡皇龍(Hsin-Tsai Liu),余天心(Chih-Yang Tseng) | |
| dc.subject.keyword | 葉綠體,蛋白運輸,雙陽性模體,葉綠體外膜機組蛋白159（Toc159）, | zh_TW |
| dc.subject.keyword | Chloroplasts,Protein import,Twin-positive motif,Translocon at the outer envelope membrane of chloroplast 159 (Toc159), | en |
| dc.relation.page | 44 | |
| dc.identifier.doi | 10.6342/NTU202101122 | |
| dc.rights.note | 同意授權(限校園內公開) | |
| dc.date.accepted | 2021-06-24 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
| 顯示於系所單位： | 分子與細胞生物學研究所 | |
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