臨界現象與膜融合的關聯性

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姓名

葛廷軒(Ting-Hsuan Ko) 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

臨界現象與膜融合的關聯性
(The correlation of critical fluctuation and membrane fusion)

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至系統瀏覽論文 (2026-8-16以後開放)

摘要(中)

研究人員發現到膜蛋白與膽固醇並非均勻分布於細胞膜，而是形成異質性結構。特定膜蛋白與膽固醇集結的區塊被稱作脂筏。鑒於目前的技術尚無法直接在細胞膜上觀測到脂筏，脂筏的形成機制仍屬未知。脂筏得以透過相分離形成是當前的主流觀點之一，但由細胞膜分離出來的仿生物膜卻無法在人體溫度下發生相分離行為。臨界現象為脂筏的形成機制開闢了新的方向。只有當系統非常接近臨界點時才會發生臨界漲落。臨界漲落會使局部的組成出現消長並形成存在時間極短的動態區塊。近期一份研究指出，相分離形成的區塊邊界對HIV病毒的膜融合有顯著影響。本研究將探討臨界漲落產生的動態區塊，即使存在時間非常短，是否同樣能促進膜融合。本實驗分別利用螢光光譜儀與小角度X光散射儀量測膜融合與脂雙層膜的結構。我們致力於發掘臨界漲落如何影響到生物膜結構，從而改變生成膜融合中間態所需的能量。膜融合中間態是掌握膜融合能否完成的關鍵。我們期待針對臨界漲落與膜融合的研究成果，能為對抗冠狀病毒提供新穎的見解。

摘要(英)

Researchers have found that cell membranes are heterogeneous, in the sense that membrane proteins and cholesterol tend to segregate and be unevenly distributed within cell membranes. Domains arising from the segregation is known as lipid rafts. The exact mechanisms for raft formation remains a debate due to the lack of direct observations of rafts in cell membranes. While phase separation observed in model membranes is one of the leading explanations for raft formation, no phase separation can be observed at physiological temperatures on the giant plasma membrane vesicles directly derived from cellular plasma membranes. Hence, critical fluctuation emerges as an alternative explanation for rafts. Critical fluctuation arises only when homogeneous membrane is extremely close to the critical point. At this point, it shows fluctuations in local composition and transiently forms segregation domains. A recent research found that phase separation, which spawns line tension on domain boundaries, had a significant effect on membrane fusion for HIV virus.
In this thesis work, we explore whether critical fluctuation also facilitates membrane fusion even though it only transiently forms domains. Fluorescent spectroscopy and small angle X ray scattering are employed to measure the membrane fusion efficiency and membrane structure, respectively. We aim to understand how critical fluctuation modulates the membrane structure and thereby varies the energy barriers for forming fusion intermediate structures, which in turn dictates the fusion efficacy. The results are expected to have broad implications for the fights against the membrane-bound coronavirus.

關鍵字(中)

★ 臨界漲落
★ 膜融合

關鍵字(英)

論文目次

摘要 i
Abstract ii
致謝 iii
目錄 iv
圖目錄 vi
表目錄 viii
第一章、緒論 1
1-1細胞膜 (Cell membrane) 1
1-2 脂筏 (Lipid raft) 2
1-3 微脂體 (Liposome) 4
1-4 相分離 (Phase separation) 5
1-5 臨界現象 (Critical phenomena) 8
1-6 膜融合 (Membrane fusion) 13
1-7 研究動機 (Motivation) 16
第二章、實驗方法 17
2-1實驗材料 17
2-1.1 磷脂質 17
2-1.2 螢光染劑 20
2-1.3 緩衝溶液 21
2-1.4 其他材料 22
2-2 樣品製備 23
2-2.1 微脂體製備 23
2-2.2 微脂體 (含有螢光染劑) 製備 24
2-3 粒徑大小與分布的測定 26
2-4 膜融合的量測 28
2-4.1 螢光光譜儀量測 30
2-5 小角度X光散射 (Small angle X-ray scattering, SAXS) 33
2-5.1 小角度X光散射數據處理 34
第三章、實驗結果 38
3-1 微脂體粒徑的大小與分布 38
3-2單層囊泡結構 42
3-3 膜融合與臨界現象 45
3-3.1 液相共存區 45
3-3.2 液態無序相 (Ld) 51
3-3.3 液態有序相 (Lo) 57
3-4 膜厚與臨界現象 63
3-4.1 液相共存區 63
3-4.2 液態無序相 (Ld) 66
3-4.3 液態有序相 (Lo) 69
第四章、討論 72
4-1 膜厚與臨界現象 72
4-2 膜融合與臨界現象 73
第五章、結論 77
文獻目錄與參考資料 78
附錄 82

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指導教授

陳儀帆(Yi-Fan Chen)

審核日期

2021-8-18

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