跳到主要內容

臺灣博碩士論文加值系統

(44.197.230.180) 您好!臺灣時間:2022/08/20 09:35
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:劉高超
研究生(外文):Kao-Chao Liu
論文名稱:鈣調素結構穩定度以及與嗅覺神經的核甘酸離子閘之鈣調素鍵結區之作用研究
論文名稱(外文):The stability of Calmodulin and the effect of Calmodulin binding site of cyclic-nucleotide-gated olfactory ion channel
指導教授:錢嘉琳
指導教授(外文):Chia-Lin Chyan
學位類別:碩士
校院名稱:國立東華大學
系所名稱:化學系
學門:自然科學學門
學類:化學學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:60
中文關鍵詞:鈣調素變性
外文關鍵詞:calmodulindenaturation
相關次數:
  • 被引用被引用:0
  • 點閱點閱:189
  • 評分評分:
  • 下載下載:8
  • 收藏至我的研究室書目清單書目收藏:0
中文摘要


鈣調素(calmodulin)有兩個區域(N-domain and C-domain),調控生物體內許多生化反應,本論文第一部份以GdnHCl (guanidine hydrochloride)及尿素(Urea)做為變性劑,以遠紫外圓二色光譜及螢光來測定野生種鈣調素及其三種點變異的穩定度。由實驗的結果中求出鈣調素與其變異種的穩定度( ),並討論鈣離子的鍵結對此四種蛋白質穩定度的影響,另外為N端的穩定度,亦由相同實驗中求得。
第二部份將要探討鈣調素與�a�d間的作用,藉由遠紫外圓二色光譜來做偵測,探討鈣調素與不同�a�d結合的結構變化以及比例(stoichiometric ratio),結果顯示,核甘酸離子閘的鈣調素鍵結區在與鈣調素鍵結後結構為α-螺旋,鍵結比例為1:1。
Abstract

Calmodulin (CaM) is a 148-residue protein which serves as the promary receptor for intracellular Ca2+. The structure of CaM contains two globular domains and a long sentral helix. In the first part of the thesis, we have performed the chemical denaturation experiments to determine the stability of wild type, three point-mutated, and the N-terminal domain of CaMs. Based on the two-state and three-state unfolding model, the free energy of unfolding (ΔGo) and the transition midpoint of CaM and its mutants have been calculated. The stability of apo and holo Y99W, Y138W was almost unchanged compared to wild type CaM. Interestingly, the stability of Y99WY1338W was decreased slightly in the holo form however, increased by about 0.5 kcal/mol compared to wild type CaM and Y99W and Y138W. The N-terminal domain of CaM is more stable than the C-domain in their apo-forms. However, upon binding to Ca2+, the C-terminal domain is more stable than the N-terminal one. In the second part of the thesis, we have investigated the structural change and the binding stoichiometric ratio of CaM with the binding domain of a nucleotide-gated ion channel. We have found that the peptide adopted α-helical structure upon binding to CaM and that the binding stoichiometric ratio is 1:1.
中文摘要 I
英文摘要 II
謝誌 III
目錄 IV
表目錄 V
圖目錄 VI
一、 文獻探討 1
二、 實驗步驟 9
三、 實驗結果 15
四、 實驗討論 21
五、 參考文獻 31
五、參考文獻
1.Lehninger Principles of Biochemistry. Third edition. Edit by David L. Nelson, Michael M. Cox, Chapter 5:Amino Acid, peptide, and Proteins, 151.
2.Gayatri Desai, Gunda Panick, Marion Zein, Roland Winter, and Catherine A. Royer (1999) J. Mol. Biol. 288, 461-475.
3.Gunda Panick, Ralf Malessa, Roland Winter, Gert Rapp, Kelly J. Frye and Catherine A. Royer (1998) J. Mol. Biol. 275, 389-402.
4.Circular Dichroism & linear Dichroism. Edit by Rodger and Nordén, Chapter 2:Circular dichroism of biomolecules, 20.
5.Chi-Wan Lee, Zhi-Ping Zhuang, Virginia M. -Y. Lee, and Hank F. Kung (2001) J. Med. Chem., 44, 2270 –2275.
6.Babu, Y. S., Bugg, C. E., Cook, W. J. (1988) J Mol Biol 204, 191-204.
7.Mark A. Wilson and Axel T. Brunger (2000) J. Mol. Biol. 301, 1237- 1256.
8.Andreas Barth, Stephen R. Martin, and Peter M. Bayley (1998) J. Biol. Chem. 273, 2174-2183.
9.JC. Lee and AM. Edelman (1994) J. Biol. Chem. 269, 2158-2164.
10.H. Mochizuki, R. Sugita, T. Ito, H. Hidaka (1993) Biochem. Biophys. Res. Commun. 197, 1595-1600.
11.Masanori Osawa, Mark B. Swindells, Jun Tanikawa,Toshiyuki Tanaka, Toshiyasu Mase, Toshio Furuya, and Mitsuhiko Ikura (1998) J. Mol. Biol. 276, 165-176.
12.Guy Vergéres, and Jeremy J. Ramsden (2000) Arch. Biochem. Biophys.378, 45-50.
13.Veronika Harmat, Zsolt Böcskei, Gábor Náray-Szabó, Imre Bata, Andrea S. Csutor, István Hermecz, Péter Arányi, Beáta Szabó, Károly Liliom, Beáta G. Vértessy, and Judit Ovádi (2000) J. Mol. Biol. 297, 747-755.
14.James. P., Vorherr, T. and Carafoli, E. (1995) Trends Biochem. Sci. 20, 38-42.
15.Patrick L. Wintrode and Peter L. Privalov (1997) J. Mol. Biol. 266, 1050-1062.
16.Bettina Elshorst, Mirko Hennig, Ernesto Carafoli (1999) Biochemistry, 38, 12320-12332.
17.Tao Yuan, Michael P. Walsh, Cindy Sutherland, Heinz Fabian, and Heins J. Vogel (1999) Biochemistry 38, 1446-1455.
18.Bettina Elshorst, Mirko Hennig, Ernesto Carafoli (1999) Biochemistry, 38, 12320-12332.
19.Monika Siedlecka, Grazyna Goch, Andrzej Ejchart, Heinrich Sticht, and Andrzej Bierzy ski (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 903-908.
20.Maria Luisa Ruiz, Barry London, and Bernado Nadal-Ginard (1996) J. Mol. Cell Cardiol 28, 1453-1461.
21.Hongy Sun, Dan Yin, Laurel A. Coffeen, Madeline A. shea, and Thomas c. Squier (2001) Biochemistry, 40, 9605-9617.
22.Laura Masino, Stephen R. Martin, and Peter M. Bayley (2000) Protein science 9, 1519-1529.
23.Protein structure. Edited by T.E. Greighton, Chapter 10:How to determine the molar absorbance coefficient of a protein.
24.Mineyuki Mizuguchi, Munehito Arai, Yue Ke, Katsutoshi Nitta, Kunihiro Kuwajima (1998) J. Mol. Biol. 283, 265-277.
25.Protein structure. Edited by T.E. Greighton, Chapter 12:Measuring the conformational stability of a protein.
26.Sorensen BR, Shea MA (1998) Biochemistry 37, 4244-4253.
27.Hiroshi Tokumitsu, Masa-aki Muramatsu, Mitsuhiko Ikura, and Ryoji Kobayashi. J.Biol.Chem. 275, 20090-20095.
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top