跳到主要內容

臺灣博碩士論文加值系統

(3.237.6.124) 您好!臺灣時間:2021/07/24 02:44
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:吳京穎
研究生(外文):Jing-Yiing Wu
論文名稱:真核核醣蛋白L17和L26與內質網之交互作用
論文名稱(外文):The Interaction between the Ribosomal Proteins and Endoplasmic Reticulum(ER):The role of large subunit ribosomal proteins L17 and L26 in making rough ER
指導教授:林茂榮林茂榮引用關係
指導教授(外文):Alan Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:真核核醣蛋白重組核醣體核醣蛋白L17核醣蛋白L26
外文關鍵詞:eukaryotic ribosomal proteinrecombinant ribosomeribosomal protein L17ribosomal protein L26
相關次數:
  • 被引用被引用:2
  • 點閱點閱:114
  • 評分評分:
  • 下載下載:6
  • 收藏至我的研究室書目清單書目收藏:0
關於真核核醣體與內質網結合的研究,目前的資訊除了用冷凍電子顯微鏡以15.4Å解像力對兩者間可能的四到五個核醣蛋白做接觸點預測外,尚缺乏直接實驗證明。本篇論文之研究目的乃探討其中人類核醣蛋白L17和L26與內質網結合的現象。依據古生菌50S的2.4Å之結構,真核細胞L26為所預測的四個接觸點其中之一,L17不在預測內但它位於新生胜肽鏈通道出口附近與其他接觸點相當靠近。在此研究中我們應用重組核醣體之技術,發現含磷酸化標的序列之L17與L26融合蛋白均會參與重組核醣體的組成並具有正常轉譯功能,同時發現此兩蛋白質之C端均暴露於核醣體外。在內質網之機制上我們分別運用in vitro及in vivo實驗設計,以純化之L17蛋白質與含重組核醣體之顆粒性內質網,利用內質網微粒漂浮分析法得到L17與L26會與內質網結合的結果。而於進一步利用交叉結合分析內質網與L17所連結之蛋白質的實驗結果,發現不與已知之組成穿膜孔道的蛋白質發生交互作用,因此推測L17可能與內質網上其他高分子蛋白連結。
The interaction between the ribosome and ER (endoplasmic reticulum) has been revealed by cryo-electron microscopy at 15.4Å resolution. The information predicts that four contact points, namely L19, L25, L26 and L35 of ribosome, provide the major attachments to the ER. So far, no direct biochemical evidence has been given in respect to the attachments. This thesis intends to investigate the binding of human ribosomal protein L17 and L26 to the ER. In this study I have demonstrated that the phosphorylation- tagged L17 and that of L26 can be assembled into recombinant ribosomes, and maintain a normal functions in translation. In addition I have detected that the using phosphorylation probing C-terminal regions of L17 and that of L26 expose on the surface of the ribosome. In test of purified ribosomal proteins binding the rough ER it shows that the L17 and L26 can bind to ER by the microsome floating assay. Furthermore, in a cross-linking experiment I have observed that L17 does not crosslink to the components of translocon protein complex, but rather associates with other high molecular weight proteins of ER. This result of this study has provided a useful information in understanding the mechanism of protein translocation.
Ban, N., Freeborn, B., Nissen, P., Penczek, P., Grassucci, R.A., Sweet, R., Frank, J., Moore, P.B., and Steitz, T.A. (1998) A 9Å resolution X-ray crytallographic map of the large ribosomal subunit. Cell 93: 1105-1115.
Ban, N., Nissen, P., Hansen, J., Moore, P., and Steitz, T.A. 2000) The complete atomic structure of the large ribosomal subunit at 2.4Å resolution. Science 289: 905-920.
Beckmann, R., Spahn, C.M., Eswar, N., Helmers, J., Renczek, P.A., Sali, A., Frank, J., and Blobel, G. (2001) Architecture of the protein-conducting channel associated with the translating 80S ribosome. Cell 107: 361-372.
Borel, A.C. and Simon, S.M. (1996) Biogenesis of polytopic membrane proteins: membrane segments assemble within translocation channels prior to membrane integration. Cell 85: 379-389.
Hedge, R.S. and Lingappa, V.R. (1996) Sequence-specific alteration of the ribosome-membrane junction exposes nascent secretory proteins to the cytosol. Cell 85: 217-228.
Johnson, A.E. and van Waes, M.A. (1999) The translocon: a dynamic gateway at the ER membrane. Ann. Rev. Cell Dev. Biol. 15: 799-842.
Kalies, K.U., Gorlich, D., and Rapapart, T.A. (1994) Binding of ribosomes to the rough endoplasmic reticulum mediated by the Sec61p-complex. J. Cell boil. 126: 925-934.
Lauring, B., Sakai, H., Kreibich, G., and Wiedmann, M. (1995) Nascent polypeptide-associated complex protein prevents mistargeting of nascent chains to the endoplasmic reticulum. Proc. Natl. Acad. Sci. USA 92: 5411-5415.
Liao, S., Lin, J., Do, H., and Johnson, A.E. (1997) Both lumenal amd cytosolic gating of the aqueous ER translocon pore are regulated from inside the ribosome during membrane protein intergration. Cell 90: 31-41.
Lütcke, H. (1995) Signal recognition particle (SRP), a ubiquitous initiator of protein translocation. Eur. J. Biochem. 228: 531-550.
Matlack, K.E., Mothes, W., and Rapoport, T.A. (1998) Protein translocation: tunnel vision. Cell 92: 381-390.
Nissen, P., Hansen, J., Ben, N., Moore, P.B., and Steitz, T.A. (2000) The structural basis of the ribosome activity in peptide bond synthesis. Science 289: 920-930.
Nomura, M. (1973) Assembly of bacterial ribosomes. Science 179: 864-873.
Ramakrishnan, V. (2002) Ribosome structure and the mechanism of translation. Cell 108: 557-572.
Ramakrishnan, V., and Moore, P.B. (2001) Atomic structures at last: the ribosome in 2000. Curr. Opin. Struct. Biol. 11: 144-154.
Rapoport, T.A., Roll, M.M., and Jungnickel, B. (1996) Approaching the mechanism of protein transport across the ER membrane. Curr. Opin. Cell Biol. 8: 499-504.
Rapoport, T.A., and Akey, C.W. (2000) The structure of ribosome-channel complexes engaged in protein translocation. Mol. Cell 6: 1219-1232.
Savitz, A.J., and Meyer, D.I. (1990) Idenfication of a ribosome receptor in the rough endoplasmic reticulum. Nature 346: 540-544.
Serganov, A., Ennifar, E., Portier, C., Ehresmann, B., and Ehresmann, C. (2002) Do mRNA and rRNA binding sites of E. coli ribosomal protein S15 share common structural determinants? J. Mol. Biol. 320: 963-978.
Solomon, E.P., Berg, L.R., Martin, D.W., and Villee, C. (1996) Biology, Fourth Edition, Saunders College.
Sommerville, J. (1986) Nucleolar structure and ribosome biogenesis. Trends Biochem. Sci. 11: 438-442.
Spahn, C.M., beckmann, R., Eswar, N., Penczek, P.A., Salli, A., Blobel, G., and Frank, J. (2001) Structure of the 80S ribosome from Saccharomyces cerevisiae- tRNA-ribosome and subunit-subunit interaction. Cell 107: 373-386.
Walter, P., and Blobel, G. (1983) Methods Enzymol. 96: 84-93.
劉宏文 (2001) 核醣蛋白質L7的胺基重複序列在內質網結合與核醣體合成中之特性. 國立陽明大學遺傳學所碩士論文.
柯俊龍 (2002) 真核生物核醣蛋白質之結構與功能:核醣蛋白質L7的細胞核運輸與核醣體重組. 國立陽明大學遺傳學所碩士論文.
王怡安 (2003) 建構重組核醣體:利用in vivo組合重組核醣體分析真核生物核醣蛋白之功能. 國立陽明大學遺傳學所碩士論文.
陳映潔 (2004) 真核核醣蛋白L35與內質網之交互作用. 國立陽明大學遺傳學所碩士論文.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top
無相關期刊