臺灣博碩士論文加值系統

中文摘要


鈣調素(calmodulin)有兩個區域(N-domain and C-domain)，調控生物體內許多生化反應，本論文第一部份以GdnHCl (guanidine hydrochloride)及尿素(Urea)做為變性劑，以遠紫外圓二色光譜及螢光來測定野生種鈣調素及其三種點變異的穩定度。由實驗的結果中求出鈣調素與其變異種的穩定度( )，並討論鈣離子的鍵結對此四種蛋白質穩定度的影響，另外為N端的穩定度，亦由相同實驗中求得。
第二部份將要探討鈣調素與�a�d間的作用，藉由遠紫外圓二色光譜來做偵測，探討鈣調素與不同�a�d結合的結構變化以及比例(stoichiometric ratio)，結果顯示，核甘酸離子閘的鈣調素鍵結區在與鈣調素鍵結後結構為α-螺旋，鍵結比例為1：1。

Abstract

Calmodulin (CaM) is a 148-residue protein which serves as the promary receptor for intracellular Ca2+. The structure of CaM contains two globular domains and a long sentral helix. In the first part of the thesis, we have performed the chemical denaturation experiments to determine the stability of wild type, three point-mutated, and the N-terminal domain of CaMs. Based on the two-state and three-state unfolding model, the free energy of unfolding (ΔGo) and the transition midpoint of CaM and its mutants have been calculated. The stability of apo and holo Y99W, Y138W was almost unchanged compared to wild type CaM. Interestingly, the stability of Y99WY1338W was decreased slightly in the holo form however, increased by about 0.5 kcal/mol compared to wild type CaM and Y99W and Y138W. The N-terminal domain of CaM is more stable than the C-domain in their apo-forms. However, upon binding to Ca2+, the C-terminal domain is more stable than the N-terminal one. In the second part of the thesis, we have investigated the structural change and the binding stoichiometric ratio of CaM with the binding domain of a nucleotide-gated ion channel. We have found that the peptide adopted α-helical structure upon binding to CaM and that the binding stoichiometric ratio is 1:1.

中文摘要 I
英文摘要 II
謝誌 III
目錄 IV
表目錄 V
圖目錄 VI
一、 文獻探討 1
二、 實驗步驟 9
三、 實驗結果 15
四、 實驗討論 21
五、 參考文獻 31

五、參考文獻
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