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標題: | 探討母雞白蛋白溶菌酶聚集現象及尋找阻礙其聚集作用之抑制劑 Investigating of Amyloid Fibril Formation of Lysozyme and Inhibitors |
作者: | Po-Han Chen 陳柏翰 |
指導教授: | 王勝仕 |
關鍵字: | 溶菌酶,類澱粉纖維,抑制劑,聚集, lysozyme,amyloid fibril,inhibitor,aggregation, |
出版年 : | 2005 |
學位: | 碩士 |
摘要: | 近年來已經發現了至少十六種以上與人類有關之蛋白質因不正常地折疊而導致稱為類澱粉纖維(amyloid fibril)之物質生成及身體上之病變。 這些蛋白質雖無相似之序列及功能;且在臨床、病理、及生物化學上,這些疾病並不直接相關且各具有其獨特之特性;然而這群疾病卻有著相同的發生機制;且對造成這群疾病之類澱粉蛋白質而言,都能夠自我組合或是自我聚集成具有獨特二級結構 β-sheet 之穩定纖維。
本研究對類蛋白質(amyloid protein)聚集狀態(aggregation state),與抑制蛋白質形成類澱粉纖維的抑制劑(melatonin、benzoquinone)效果,與蛋白質周圍的環境因子(如:環境溫度、溶劑組成)間之關係進行探討。為了尋求類蛋白質與抑制劑間一般化交互作用機制,研究中採用雞蛋白溶菌?蛋白質進行上述目的之探討。期望以一些生物物理(biophysical)方法及光譜技術,例如:ThT 螢光、Congo red 鍵結與濁度測試,並配合動力學進行數據分析討論。研究發現,鹽酸溶液(pH 2.0)可以成功誘導出類澱粉纖維;且具鹽類之鹽酸溶液(pH 2.0)比起不具鹽類之鹽酸溶液(pH 2.0)所誘導之類澱粉纖維於時間上更加快速,數量上也更加多。此篇研究選用退黑激素與苯昆來研究是否會阻礙蛋白質自我聚集,結果指出苯昆比起退黑激素具有良好的抑制作用,對於不同溫度誘導類澱粉纖維測試,低溫下誘導之類澱粉纖維所需時間較高溫慢。此研究之成果將可以用於解釋抑制劑對於蛋白質的交互作用。 More than sixteen different human proteins can fold abnormally resulting in the formation of amyloid fibrils and accompanying pathologies. These proteins have little sequence homology; however, they can polymerize into stable fibrils with a characteristic cross β-sheet structure. Although amyloid diseases have been the center of intense research, the role of amyloid proteins and the associated cytotoxicity mechanisms that mediate biological responses remain largely unknown. This research, with hen egg white lysozymes utilized as a model system, was set out to study the mechanism(s) of aggregation/fibril formation. First part of the research not only sought out the condition(s) of forming amyloid fibrils but also investigated the relationship between aggregation/fibril formation and environmental factors such as ambient temperature and solvent conditions. Secondly, this work was aimed at examining the inhibitory action of two potential compounds, namely melatonin and benzoquinone, against the aggregation/fibril formation of lysozyme. It was demonstrated that the fibrillar form of hen egg while lysoyzme was able to be produced under the acidic condition (pH 2.0). In addition, the elevated rate of amyloid fibril formation was observed upon salt addition. Our study then showed that benzoquinone exhibited a better anti-aggregating action than melatonin. Finally, in regards to the temperature effect, the lysozymes formed fibrils faster at 70°C in comparison with 37°C and 50°C. We believe that the outcome from this work may contribute to a better understanding of how amyloid proteins polymerize and further facilitate the development of possible strategies to prevent lysozyme or other amyloid protein-associated diseases. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/38311 |
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顯示於系所單位: | 化學工程學系 |
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