自發曲率、金屬離子吸附以及微脂體膜融合效率三者間之相關性探討

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姓名

樊子安(Zih-an Fan) 查詢紙本館藏

畢業系所

化學工程與材料工程學系

論文名稱

自發曲率、金屬離子吸附以及微脂體膜融合效率三者間之相關性探討
(The Correlation among Spontaneous Curvature, Metal Ions Binding and Membrane Fusion of Liposomes)

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摘要(中)

本研究旨在探討monolayer spontaneous curvature、金屬離子binding以及膜融合三者之間的關係。膜融合是在細胞的生命活動中一個重要且常見的過程，例如：包有神經傳導物質的囊泡藉由鈣離子觸發，與神經細胞的細胞膜進行膜融合並將神經傳導釋出。我們將dioleoyl phosphatidylethanolamine/dioleoyl phosphatidylcholine/4 mol% dioleoyl phosphatidic acid脂質混合物製備成可以模擬生物系統的unilamellar vesicle以及為了量測spontaneous curvature的dispersion兩種樣品。影響膜融合的因素包括monolayer spontaneous curvature以及金屬離子binding。Monolayer spontaneous curvature（C0）是一種彈性性質，它量化脂質分子形成lamellar phase或是nonlamellar phase的趨勢，而nonlamellar phase的形成則被認為是在膜融合中一個重要的過程。從我們的實驗結果發現，膜融合的效率隨著spontaneous curvature的增加而增加。當金屬離子binding在脂質膜時會影響脂質膜的穩定性以及性質。從我們的實驗結果發現，spontaneous curvature和金屬離子binding皆會使脂質膜的厚度增加，但是，膜厚的變化並不足以影響膜融合；DOPE與DOPC頭基的不同造成金屬離子binding對spontaneous curvature的影響程度不同。我們也發現，金屬離子binding影響intra-monolayer與inter-monolayer，進而改變spontaneous curvature與脂質的自組裝結構，最後影響膜融合的效率。

摘要(英)

In the present work, we investigate the correlation among spontaneous curvature, metal ions binding and membrane fusion of liposomes. Membrane fusion is an important process of many cellular events, such as neuronal signaling. In order to simulate the biological system, we use DOPE/DOPC/4 mol% DOPA lipid mixture to prepare the unilamellar vesicle. We also prepare lipid dispersion for the measurement of monolayer spontaneous curvature. There are many factors that can affect membrane fusion, including the monolayer spontaneous curvature and metal ions binding. Monolayer spontaneous curvature (C0) quantifies the tendency of forming lamellar phase or nonlamellar phase. Forming nonlamellar structure is considered to be associated with membrane fusion. When the spontaneous curvature become more negative, membrane fusion efficiency will increase. The binding of metal ions to biological membranes affects the stability and the properties of lipid bilayer, is associated with membrane fusion. Both spontaneous curvature and metal ions binding induce lipid membrane thicker. However, the difference of membrane thickness is too small to affect membrane fusion. The influence of metal ions binding to DOPE and DOPC spontaneous curvature is different, due to the difference of lipid headgroup composition. The binding of metal ions to lipid membrane affects lipid intra-monolayer and lipid inter-monolayer, changing the spontaneous curvature and liposome structure, and then, membrane fusion efficiency will be impacted.

關鍵字(中)

★ 自發曲率
★ 金屬離子結合
★ 微脂體
★ 膜融合

關鍵字(英)

★ spontaneous curvature
★ metal ions binding
★ liposome
★ membrane fusion

論文目次

摘要 i
Abstract iii
致謝 iv
目錄 v
圖目錄 vii
第一章、緒論 1
1-1. 脂質（Lipid） 1
1-1-1. 磷脂質（Phospholipid） 1
1-1-2. 細胞膜（Cell membrane） 4
1-2. 彈性性質（Elastic property） 5
1-2-1. 自發曲率（Spontaneous curvature） 5
1-3. 微脂體（Liposome） 8
1-4. 膜融合（Membrane fusion） 9
1-4-1. 膜融合的機制（Mechanism of membrane fusion） 10
1-4-2. SNARE蛋白 11
1-4-3. 聚乙二醇（Poly(ethylene) glycol，PEG）誘導膜融合 12
1-5. Spontaneous curvature與膜融合（Membrane fusion）之間的關係 13
1-6. 金屬離子與膜融合（Membrane fusion）之間的關係 16
1-7. 研究動機 19
第二章、實驗方法 20
2-1. 實驗材料 20
2-2. 實驗儀器 24
2-3. 樣品製備 25
2-3-1. 微脂體的製備方法 25
2-3-2. 微脂體（含有螢光染劑）的製備方法 27
2-3-3. Dispersion（hexagonal phase）的製備方法 31
2-4. 奈米粒徑分佈的量測 34
2-5. 膜融合的測定 35
2-6. 螢光光譜儀（Spectrofluorometer）量測 39
2-6-1. 膜融合的量測 39
2-7. 小角度X光散射（Small-Angle X-ray Scattering） 44
2-8. 數據處理（Data process） 45
2-8-1. 微脂體（Liposome）數據處理 46
2-8-2. Hexagonal phase數據處理 48
第三章、結果 53
3-1. 微脂體（Liposome） 53
3-1-1. 單層囊泡（Unilamellar vesicle）的粒徑大小與均一度
（Monodispersity） 53
3-1-2. 單層囊泡（Unilamellar vesicle）的結構 56
3-1-3. 脂質膜的膜厚（Membrane thickness）與Spontaneous curvature
之間的關係 57
3-2. Spontaneous curvature與膜融合之間的關係 58
3-2-1. DOPE/DOPC的Spontaneous curvature 58
3-2-2. Spontaneous curvature對膜融合之影響 60
3-3. 金屬離子Binding與膜融合之關聯性 62
3-3-1. 金屬離子Binding對微脂體（Liposome）結構之影響 62
3-3-2. 金屬離子Binding對脂質膜厚度（Membrane thickness）
之影響 66
3-3-3. 金屬離子Binding對膜融合之影響 68
3-4. 金屬離子Binding與Spontaneous curvature之關係 70
第四章、討論 75
4-1. 膜厚與Spontaneous curvature、金屬離子Binding 之關聯性 75
4-2. Spontaneous curvature對膜融合之影響 76
4-3. 金屬離子Binding與微脂體結構、膜融合之關聯性 78
4-4. 金屬離子Binding與DOPE、DOPC Spontaneous curvature之關聯性 80
4-5. Spontaneous curvature、金屬離子Binding與膜融合之關聯性 82
第五章、結論 91
參考文獻 94

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指導教授

陳儀帆(Yi-fan Chen)

審核日期

2015-7-29

推文