臺灣博碩士論文加值系統

Biological membranes mostly composed of phospholipid molecules and proteins form a barrier between intra and extra-cellular environments and be able to control the material enters and exit the cell. Both in vivo and in vitro membrane destabilization can usually be induced by external agents such as bio molecules (i.e., proteins and peptides).
Fast field cycling nuclear magnetic resonance relaxometry can provide valuable information because of it is sensitivity to dynamic process that occur over a broad time scale. Recorded the data for the large unilamellar liposome which compose of POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol) and POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) with addition of bacitracin and Ca2+. By analysis of the data recorded, the result shown that the influence of the bacitracin depends on its concentration. Bacitracin mainly affected the order fluctuation of the membrane, diffusional motion and rotational correlation time liposome. Moreover, the P/L ratio with higher concentration caused weaker surface charge and less shielding effect of the liposome.

Key word : NMRD, bacitracin, phospholipid membrane.

Contents
Abstract……………………………………………………………………...………..…i
Contents ………………………………………………………………………..…….…ii
List of Figures………………………………………………………………….……….iv

List of Tables ………………………………………………………………….……….v

Abbreviation list ……………………………………………………………….……….vi

Chapter 1 Introduction ……………………………………………………………….....1
1-1 Lipid bilayer ………………………………………………………………….……..1

1-2 Liposome …………………………………………………………………………...2

1-2-1Liposome Carriers of Different Types ………………………………….…….5

Chapter 2 Theory and Studies ……………………………………………………….….8
2-1 Preparation of Lipid For Hydration ………………………………………………..8
2-2-1 Antimicrobial Peptides (AMPs) ……………………………………………...9
2-1-2 Structure and Major Activities of AMPs …………………………….……….11
2-1-3 Bacitracin ……………………………………………………………….……14
2-1-4 Bloch Equation ………………………………………………………….…...17
2-1-5 Relaxation ……………………………………………………………….…...18
2-1-5-1 Spin Lattice Relaxation Mechanism ……………………………………….18
2-1-5-2 Spin-Spin Relaxation Time Measurement.……………………….…..……21
2-1-5-2-1 Relaxation Models For Biomembranes …………………….………..….21
2-1-6 Flourescence Spectroscopy………………………………………..………....23

2-1-7 Dynamic Light Scattering (DLS) ……………………………………….…....27
2-1-8 CD (Spectrum) ……………………………………………….…………..…..28
Chapter 3 Experiment …………………………………………………….………….….33
3-1 Experimental Chemicals ……………………………………….…………………...33
3-2 Method ……………………………………………………….……………………..35
3-2-1 Sample Preparation………………………………………………….…...35

3-2-2 Experimental Conditions ………………..…………………………..…..35
3-2-3 Molecular Calculation of Phospholipid on The Surface………………..36
3-3-2 Fluorescence Spectrometry…………………………………..…………..37
3-3 Experimental Section ……………………………………….……………….…...…37
3-3-1 CD (Circular Dichroism)…………………………………………………….…..38

3-4 Nuclear Magnetic Resonance ………………………………..……………..….…..39
3-4-1 FFC- NMR ………………………………………………………………….....….39
Chapter 4 Experimental result and discussion ……………………………….…..…….43
4-1 FFC-NMR result ……………………………………………………….…...….……43
4-1-1Comparison of NMRD profiles in each sample ……………..……..…….43
4-1-2 Order fluctuation of liposome …………………….………….………….47
4-1-3 Translational diffusion of lipids molecule ………..……….……….…….48
4-1-4 Rotational correlation time of liposome …………..………..…….………49
4-2 Fluorescence Spectrometry……………………………….…………….…….…....51
4-2-1 Calcein Leakage Assay Result ……………………….………………….……..…51
4-3 CD (Spectrum) …………………………………………………….………….….….52
4-3-1 Determination of Secondary Structure of Bacitracin ………...…….……..…..52
Chapter 5 Summary and prospects …………………………….………………….........54
5-1 Summary . ………………………………………….………….………….…….…...54
5-2 Prospects ………………………………………………………….………………....54
References ……………………………………………………………….….…………..55

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