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研究生:謝怡慧
論文名稱:結合傅氏紅外線光譜儀及電腦模擬計算研究眼鏡蛇磷脂水解酵素A2與細胞膜之間的作用
論文名稱(外文):Peripheral binding mode of cobra phospholipase A2 on phospholipid membranes as studied by combined FTIR and computer simulation approach
指導教授:吳文桂
學位類別:碩士
校院名稱:國立清華大學
系所名稱:生物資訊與結構生物研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
論文頁數:66
中文關鍵詞:衰減全反射磷脂水解酵素A2傅氏轉換紅外線光譜多層膜二級結構
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台灣眼鏡蛇分泌的毒素主要可以分成三大種;神經毒素(Neurotoxin)、磷脂水解酵素A2(Phospholipase A2)、心臟毒素(Cardiotoxin)。此三種毒液除了神經毒素具有特殊蛋白質受體外,磷脂水解酵素A2及心臟毒素被認為毒性來源及主要的作用位置是細胞膜磷脂質。本論文主要針對磷脂水解酵素A2與細胞膜作用為主要研究目標。

磷脂水解酵素A2分成好幾類,本論文中使用的台灣眼鏡蛇磷脂水解酵素A2主要是分泌性的磷脂水解酵素A2、GroupⅠA,分子量主要是13~14KD,二極結構大部分是α螺旋,本論文主要是利用傅氏紅外線光譜儀及電腦計算來得出台灣眼鏡蛇磷脂水解酵素A2在人工細胞膜上的結合角度,所以的結論為:

(一) 利用FTIR及電腦計算所得出的結果,台灣眼鏡蛇磷脂水解酵素A2在膜上是以α螺旋接近平躺的方式結合,且此結果與中研院院士蕭傳鐙老師利用SDS與磷脂水解酵素複合物所解出的結晶結構相似,兩相結果互相映證。

(二) 用FTIR及電腦計算所得出結果,蜜蜂磷脂水解酵素A2在膜上的結合角度,與之前利用EPR方法得出的結構也吻合。

繼台灣眼鏡蛇心臟毒素A3利用傅氏紅外線光譜儀解出在膜上的結合角度後,又再次利用了此技術解出磷脂水解酵素A2與膜的結合角度,重要的是心臟毒素A3的二極結構大部分是β摺版,而磷脂水解酵素A2的二極結構大部分是α螺旋,在兩種不同結構中,可以利用相同的技術解出結果,所以在未來生物分子之間研究上的應用,傅氏紅外線光譜儀可以更廣泛的被應用。
Cobra phospholipase A2, a α-helix acidic polypeptide, is known to hydrolyze membrane and involve some cell signaling pathways. Herein, we demonstrate that the major cobra phospholipase A2 ( Naja atra ), By the combined polarized attenuated total reflection infrared spectroscopy and computer simulation studies, Naja atra phospholipase A2 is shown to peripherally bind to multilayers in a similar edgewise manner with a tilted angle of 160° between theα-helix plane of the Cobra phospholipase A2 molecule and the normal of the membrane surface.
Qualitatively speaking, theα-helix plane of the Cobra phospholipase A2 molecule with (90°,60°) orientation appears to be more perpendicular to the membrane surface than that with (160°, 60°) orientation.
第一篇 緒論…………………………………… 1

第一節 蛇毒簡介……………………………………………………… 1
第二節 PLA2的介紹…………………………………………………… 5
第三節 細胞膜簡介…………………………………………………… 10
附圖……………………………………………………………………… 13

第二篇 核心實驗技術簡介…………………… 20

第一節 何謂傅氏轉換紅外線光譜…………………………………… 20
第二節 生物分子之紅外線吸收光譜帶……………………………… 21
第三節 霍氏自我去迴旋……………………………………………… 23
第四節 衰減全反射(ATR)………………………………………… 25
附圖……………………………………………………………………… 30

第三篇 實驗結果……………………………… 42

第一節 前言…………………………………………………………… 42
第二節 實驗材料及技術……………………………………………… 43
第三節 結果…………………………………………………………… 47
第四節 討論…………………………………………………………… 50
附圖……………………………………………………………………… 52

第四篇 參考資料……………………………… 60
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