本文研究蜂毒蛋白(melittin)與飽和鏈脂膜之作用。利用circular dichroism (CD)與isothermal titration calorimetry (ITC)測量蜂毒蛋白吸附飽和鏈脂膜的自由能，焓及熵，以熱力學探討吸附過程。
實驗結果顯示蜂毒蛋白吸附飽和鏈脂膜，與吸附未飽和鏈脂膜相同，皆為放熱反應，可知吸附為一個焓主導的過程，但吸附飽和鏈脂膜有異常放熱，推測蜂毒蛋白在飽和鏈脂膜上有聚集，這結果和文獻中利用螢光能量轉移技術(fluorescence energy transfer)的實驗相符。

This thesis is to study the binding of melittin to saturated lipid membrane. We use circular dichroism (CD) and isothermal titration calorimetry (ITC) to measure the binding free energy, the binding enthalpy and the binding entropy for understanding the thermodynamics of binding process.
The result reveals that, as the same as to the unsaturated lipid membrane, the melittin binding to the saturated lipid membrane is an exothermic reaction, indicating an enthalpy -driving process. However, there is an abnormal heat release for melittin binding to saturated lipid membrane, implying that melittin peptides are aggregated on membrane surface. This result is agreed with the measurement using fluorescence energy transfer technique.

[1]. D. Hultmark, H. Steiner, T. Rasmuson and H. G. Boman (1980). Eur. J. Biochem. 106:7.
[2]. M. Zasloff (1987) Proc. Natl. Acad. Sci. U.S.A. 84:5449.
[3]. Jake Bello, Helene R. Bello, and Edward Granados (1982). “ Conformation and Aggregation of Melittin: Dependence on pH and Concentration”,Biochemistry, 21:461-465
[4]. 歐陽鐘汕和劉寄星(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)。
[5]. H. W. Haung (2000). ’Action of antimicrobial peptides:two-state model’,Biochemistry, 39:8347-8352
[6]. N. K. Subbarao, R. I. MacDonald, K. Takeshita, and R. C. MacDonald (1991).‘Characteristics of spectrin-induced leakage of extruded, phosphatidylserine vesicles.’, Biochim. Biophys. Acta. 1063:147-54
[7]. R. C. MacDonald, R. I. MacDonal, B. P. Menco, K. Takeshita, N. K. Subbarao, and L. R. Hu (1991) .’Small-volume extrusion apparatus for preparation of large, unilamellar vesicles’, Biochim. Biophys. Acta. 1061:297-303
[8]. 林慶豪(2005).’抗菌小蛋白吸附脂膜的能量測量’/中央大學/物理學系/93年度/碩士論文
[9]. 洪玉珊(2007).’抗菌小蛋白吸附脂質微胞之多體效應’/中央大學/物理學系/95年度/碩士論文
[10]. Lange’s Handbook of Chemistry, 14th Edition, Dean, J.A., Ed. (1992). McGraw-Hill, Inc., New York.
[11]. Handbook of Chemistry and Physics, 56th Edition, Weast, R.C., Ed. (1975). CRC Press, Cleveland
[12]. Gill, S.C. and von Hippel, P.H. (1989).‘Calculation of protein extinction coefficients from amino acid sequence data.’,Anal. Biochem. 182:319-26
[13]. Pace, C.N., et al. (1995). ‘How to measure and predict the molar absorption coefficient of a protein.’ Protein Sci. 4:2411-23
[14]. Practical Handbook of Biochemistry and Molecular Biology, Fasman, D.G., Ed. (1992). CRC Press, Boston
[15]. Marilyn Emerson Holtzer, Alfred Holtzer (1992).“ -helix to random coil transitions: Determination of peptide concentration from the CD at the Isodichroic point”,Biopolymers. Vol.32, Issue 32:1675–1677
[16]. Horst Vogel, Fritz Jahnig (1986). ”The structure of melittin in membranes”,Biophysical journal. 50:573-582
[17]. William Wilcox, Divid Eisenberg (1992).“ Thermodynamics of melittin tetramerization determined by circular dichroism and implications for protein folding”,Protein Sci. 1: 641-653
[18]. Edgar John, Fritz Jahnig (1991).“Aggregation state of melittin in lipid vesicle membranes”,Biophysical journal. 60：319-328
[19]. D. Allende, S. A. Simon, and T. J. McIntosh (2005). “Melittin-Induced Bilayer Leakage Depends on Lipid Material Properties:Evidence for Toroidal Pores”. Biophys. J. 88:1828-1837
[20]. Alexey S. Ladokhin, Stephen H. white (1999).“Folding of Amphipathic α-helices on Membrances:Energetics of Helix Formation by Melittin”. J. Mol. Biol. 285:1363-1369