脂質組成成分對細胞膜物理性質與生物功能的影響

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臧冠遇(Kuan-yu Tsang) 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

脂質組成成分對細胞膜物理性質與生物功能的影響
(THE INFLUENCES OF LIPID COMPOSITION ON THE PHYSICAL PROPERTIES AND BIOLOGICAL FUNCTION OF THE CELL MEMBRANE)

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摘要(中)

細胞膜在生物細胞中扮演著極其重要的角色。一般來說在細胞膜中磷脂質為主要成分之一，而磷脂質種類又相當複雜包含不同頭基與雙鍵數。在本篇論文中我們有系統地比較擁有不同頭基與不同雙鍵數的脂質分子與磷脂質膜中的比例，藉由X-光繞射實驗來獲得不同種類脂質分子的靜態物理性質(如:彈性性質)，以及利用電子自旋共振光譜來量測不同種類脂質分子的動態物理性質(如:脂質分子尾端碳氫長鏈的動態資訊)，最後利用螢光光譜來探討脂質分子組成的改變對細胞膜生物功能性上的影響(如:細胞膜融合)，其目的就是為了找出影響生物過程背後的主要機制。
不同頭基(phosphatidylcholine,phosphatidylethanolamine)的脂質分子對自發曲率值與彎曲係數有很大的影響，但對脂質分子尾端碳氫長鏈的動態速率沒有明顯可觀察到的關聯性，此外隨著增加PE頭基脂質分子的數量可以明顯地增加膜融合率。相同頭基的脂質分子隨著雙鍵數的增加自發曲率並沒有明顯地改變，但會明顯地增加脂質分子的動態速率且降低彎曲係數，且膜融合率會明顯下降。
藉由本次實驗的結果，我們找出了不同種類的脂質分子(包含不同頭基與雙鍵數)對細胞膜的物理性質與生物功能有所影響。改變不同脂質頭基與雙鍵數會對膜融合率有顯著的差異，所以我們推測有兩種機制可以改變細胞膜的膜融合率。細胞膜可以透過改變彈性能量與脂質分子的動態速率促進或抑制細胞膜膜融合的發生，而改變方法可藉由不同頭基或雙鍵數的脂質分子來完成，或許這可以解釋為何在生物細胞膜中需要擁有不同頭基與雙鍵數脂質分子的原因。

摘要(英)

Cell membranes play very important roles for the cell. In general, phospholipid is one of the major constituents of a cell membrane, whereas cell membranes contain many different kinds of phospholipids with different kinds of headgroups and degrees of unsaturation. In this report, we systematically compare different lipid molecules which have different headgroups and different degrees of unsaturation. We used X-ray diffraction to determine the static physical properties (such as elastic properties) of different kinds of lipid molecules, and used electron spin resonance spectroscopy to determine the dynamic physical properties (such as the dynamics of lipid hydrocarbon chains) of different kinds of lipid molecules. Finally, we used fluorescence spectrophotometer to investigate whether different lipid compositions will affect the biological function (such as membrane fusion) of lipid membranes. We want to find out the relationship between composition (including different headgroups and different degrees of unsaturation) and membrane physical property, and whether these properties will effect biological function. And we also want to know the mechanism underlying this correlation.
The difference in headgroup (phosphatidylcholine(PC), phosphatidylethanolamine(PE)) has great influence on the spontaneous curvature and bending modulus, but it seems to have almost no effect on the dynamic of lipid hydrocarbon chain. And the membrane fusion rate increases with the amount of PE headgroup. Changing the degrees of unsaturation for both PE and PC headgroups has weak influence on the spontaneous curvature, but it would increase the dynamics of lipid hydrocarbon chain and decrease the bending modulus. And the membrane fusion rate was significantly decreased with increasing the degrees of unsaturation.
Consequently, we found out the relationships among membrane composition, membrane physical property and biological function. Changing headgroups and degrees of unsaturation has a significantly different effect on membrane fusion rate. We speculated that there are two mechanisms which can change the fusion rate of the cell membrane. We can promote or inhibit the membrane fusion rate by changing membrane elastic energy and the dynamics of lipid molecular. This may be one of the reasons why cell membranes contain many different kinds of lipids.

關鍵字(中)

★ 磷脂質
★ 自發曲率
★ 彎曲係數
★ 膜融合

關鍵字(英)

★ phospholipid
★ spontaneous curvature
★ bending modulus
★ membrane fusion

論文目次

摘要....................................................I
Abstract..............................................III
致謝....................................................V
目錄...................................................VI
圖目錄...............................................VIII
表目錄..................................................X
第一章緒論.............................................1
1-1 細胞膜..............................................1
1-2 磷脂質..............................................3
1-3 細胞膜的彈性性質.....................................8
1-4 細胞膜的流動性......................................16
1-5 細胞膜融合..........................................19
1-6 實驗目的............................................22
第二章實驗材料與方法....................................23
2-1 實驗材料............................................23
2-2 樣品製備............................................25
2-2-1 製備lipid dispersion solution-X-ray 繞射實驗......25
2-2-2 製備微脂體-電子自旋共振光譜儀與螢光光譜儀............27
2-3 實驗儀器............................................31
2-3-1 X光繞射實驗.......................................31
2-3-2電子自旋共振光譜儀..................................31
2-3-3螢光光譜儀.........................................31
2-4 脂質分子的靜態物理性質-X光繞射實驗....................32
2-4-1 X光繞射原理.......................................32
2-4-2 數據處理與分析....................................33
2-5 脂質分子的動態物理性質-電子自旋共振光譜儀..............35
2-5-1電子自旋共振光譜原理................................35
2-5-2數據處理與分析.....................................36
2-6 膜融合實驗-螢光光譜儀................................38
2-6-1 螢光光譜原理......................................38
2-6-2數據處理與分析.....................................39
第三章結果.............................................40
3-1 自發曲率............................................40
3-1-1 不同頭基對自發曲率影響.............................51
3-1-2 不同雙鍵數對自發曲率影響............................52
3-2彎曲係數.............................................54
3-2-1 不同頭基對彎曲係數影響.............................58
3-2-2 不同雙鍵數對彎曲係數影響...........................60
3-3轉動相關時間.........................................62
3-3-1 不同頭基對轉動相關時間影響..........................68
3-3-2 不同雙鍵數對轉動相關時間影響........................70
3-4 膜融合率............................................72
3-4-1 不同頭基對膜融合率影響.............................74
3-4-2 不同雙鍵數對膜融合率影響...........................75
第四章討論.............................................76
4-1 不同頭基與雙鍵數對自發曲率影響........................76
4-2 脂質分子的動態速率與彈性性質的關係.....................78
4-3 細胞膜的物理性質與生物功能的影響......................80
第五章結論.............................................87
參考文獻................................................89

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指導教授

陳儀帆(Yi-fan Chen)

審核日期

2015-7-29

推文