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研究生: 陳夙容
Su-Jung Chen
論文名稱: 以螢光染料方法研究脂質微胞的滲漏
Leakage of lipid vesicle studied by dye fluorescence method
指導教授: 陳方玉
Fang-Yu Chen
口試委員:
學位類別: 碩士
Master
系所名稱: 理學院 - 物理學系
Department of Physics
畢業學年度: 95
語文別: 中文
論文頁數: 24
中文關鍵詞: 滲漏螢光染料方法脂質微胞
外文關鍵詞: leakage, vesicle, fluorescence method
相關次數: 點閱:17下載:0
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  • 抗菌蛋白的殺菌機制為近年來廣為研究的課題。本實驗以DOPC脂質分子所製備的微胞,內含高濃度螢光染劑Calcein,並以蜂毒蛋白melittin在微胞上打洞造成螢光染劑的滲漏,來研究蜂毒蛋白與脂膜的作用。
    實驗結果顯示: 滲漏率隨著melittin的濃度而線性增加,當melittin與脂膜分子比率達到6/1000則有100%滲漏。推斷是蜂毒蛋白吸附脂膜時造成瞬間的裂縫而使滲漏率線性增加; 推斷有100%滲漏,是因蜂毒蛋白在膜上形成孔洞所致。


    The mechanism of antimicrobial peptide has been extensively studied. In this thesis, we measured the melittin-inducing leakage of calcein from DOPC vesicles to study the interaction between melittin and lipid membrane.
    The result revealed that the leakage increases in linear with the melittin concentration, and reaches to 100% leakage as the molar ratio of melittin to lipids becomes larger than 6/1000. The linear increase is attributed to the transient disruption upon melittin binding to the membrane, whereas 100% leakage is due to the pore formation induced by high concentration peptides.

    摘 要 i Abstract ii 誌 謝 iii 目錄 iv 圖目錄 v 第一章 簡介 1-1細胞膜與脂質分子的簡介 1 1-2 抗菌蛋白的簡介 3 1-3 研究動機與目的 4 第二章 實驗樣品製備 2-1 微胞的製備 6 2-2 膠體過濾法 7 2-3 脂質分子濃度測量 9 第三章 實驗方法與結果 3-1 螢光滲漏實驗的測試 13 3-2 Meilttin與脂膜作用的螢光滲漏實驗 18 第四章 討論 22 參考資料 23

    [1].歐陽鍾汕和劉寄星(1995) ”從肥皂泡到液晶生物膜”,新世紀物理研習叢書9 ,牛頓出版(台灣)
    [2].B. Alebert, et al. (1994) “molecular biology of the cell ”3th edition, Garland publishing(New York & London)
    [3].洪偉清 (2000)“脂膜的X-光片層繞射”,國立中央大學/物理與
    天文研究所/88 年度/博士論文
    [4].H. W. Huang (2000)“Action of antimicrobial peptides:
    Two-state model ” Biochemistry 39:8347-8352.
    [5].F. Y. Chen, M. T. Lee, and H. W. Huang (2002)“Sigmoidal Concentration Dependence of Antimicrobial Peptide Activities: A Case Study on Alamethicin ”Biophys.J.82:3751–3758
    [6].F. Y. Chen, M. T. Lee, and H. W. Huang (2003)“Evidence for Membrane Thinning Effect as the Mechanism for Peptide-Induced Pore Formation”Biophys.J.84:3751–3758
    [7].T. Wieprecht, O. Apostolov, M. Beyermann, and J. Seelig (2000)“membrane binding and pore formation of the antibacterial peptide PGLa :thermodynamic and mechanistic aspects Biochemistry 39:442-452
    [9].Toni Benachir, Michel Lafleur (1995) ”Study of vesicle leakage induced by melittin”Biochimica et Biophysica Acta 1235 : 452-460
    [10].Toni Benachir, Martine Monette , Julie Grenier, Michel Lafleur (1997) “Melittin-induced leakage from phosphatidylcholine vesicles is modulated by cholesterol: a property used for membrane targeting” Eur.J.Biochem.25:201-210
    [11].H. Raghuraman and Amitabha Chattopadhyay(2004) “Interaction of Melittin with Membrane Cholesterol: A Fluorescence Approach ”Biophysical Journal 87:2419-2432

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