系統識別號 | U0002-2507201414453100 |
---|---|
DOI | 10.6846/TKU.2014.01037 |
論文名稱(中文) | C末端殘基如何改變兩親性螺旋摺疊:CD和NMR的研究 |
論文名稱(英文) | How C-terminal residue alters the helix folding of an amphipathic antimicrobial peptide: CD and NMR study |
第三語言論文名稱 | |
校院名稱 | 淡江大學 |
系所名稱(中文) | 化學學系碩士班 |
系所名稱(英文) | Department of Chemistry |
外國學位學校名稱 | |
外國學位學院名稱 | |
外國學位研究所名稱 | |
學年度 | 102 |
學期 | 2 |
出版年 | 103 |
研究生(中文) | 曾雅琳 |
研究生(英文) | Ya-Lin Tseng |
學號 | 601160061 |
學位類別 | 碩士 |
語言別 | 繁體中文 |
第二語言別 | |
口試日期 | 2014-07-10 |
論文頁數 | 170頁 |
口試委員 |
指導教授
-
李長欣(cshlee@mail.tku.edu.tw)
委員 - 李長欣 委員 - 鄧金培(jpdeng@mail.tku.edu.tw) 委員 - 錢嘉琳(chyan@mail.ndhu.edu.tw) |
關鍵字(中) |
C末端 兩親性 摺疊 CD NMR |
關鍵字(英) |
Folding antimicrobial peptide C-terminal CD NMR |
第三語言關鍵字 | |
學科別分類 | |
中文摘要 |
如何從蛋白質序列摺疊成其獨特的三維結構,一直是重要的研究課題之一。在其他文獻報告指出,蛋白質的C末端殘基會影響蛋白質的摺疊行為。由於蛋白質的胺基酸數目較多,實驗的複雜性較高,為了降低實驗的複雜性,此論文中選用胺基酸數目較少的MP-B胜肽(14個胺基酸)及其衍生物,將其溶於30% TFE的水溶液中,溫度為310 K,利用CD和NMR進行研究,觀察胜肽的C末端殘基對於蛋白質摺疊行為的影響。 從CD實驗中說明,MP-B及其衍生物在30% TFE溶液中會形成α-helix結構。利用不同溫度的DOSY實驗觀察MP-B及其衍生物的疏水性與寡聚體之間的關係,並討論寡聚狀態的熱穩定性。從抗菌活性實驗的結果發現MP-B及其衍生物的抗菌效果為MP-B > MP-BL15 > MP-B1-13。將MP-B及其衍生物的NOE距離限制放入結構計算中,統計每條胜肽的100個螺旋摺疊結構,並探討C末端對於摺疊結構熱穩定性的重要性。 統合所有的實驗結果,當胜肽C末端無序的殘基無法提供足夠的熵時,會以螺旋N端展開的方式來增加熵。胜肽具有較穩定的結構和寡聚狀態時,會有較佳的抗菌活性,因此,如欲設計一個具有抗菌活性的胜肽,其C末端殘基的作用具有很關鍵的影響。 |
英文摘要 |
One of the most interesting questions in biophysics is how protein sequences determine their unique three-dimensional structure. Previous studies reported that the residues at C-terminal can have effects on the protein folding behaviors. Since a large protein is highly complex, for simplicity, in this study we use MP-B (14 amino acids) and its analogues in 30% TFE-d3 aqueous solution as a model system. By using circular dichroism (CD) and NMR methods, we can get insight into how the C-terminal segment may influence the folding behavior of proteins. Spectra of CD indicated that our peptides may form α-helical conformations in 30%TFE. Oligomerization and thermal stability of peptides were investigated by performing diffusional experiments with variable temperatures. The antimicrobial activity of MP-B1-13, MP-B and MP-BL15, is ordered as MP-B > MP-BL15 > MP-B1-13. 100 of structures of different kind foldings were calculated with NMR data to investigate the effects of C-terminal segment on folding. Our results suggested that while the disordered residues at C-terminal can’t provide enough motional entropy, the helical N-terminal may be partially unfolded to increase the entropy. When peptides have more stable structure and oligomeric state, they may have greater antimicrobial activity. The results implied the disordered C-terminal residues may play a critical role for the structure and activity of a peptide. The knowledge is especially important while a peptide segment is designed or truncated for studies of structure and function. |
第三語言摘要 | |
論文目次 |
目錄 目錄 .......................................................................................................................................... I 圖目錄 .................................................................................................................................... V 表目錄 ................................................................................................................................XIV 縮寫表.................................................................................................................. XVIII 第一章 緒論 .............................................................................................................. 1 1.1 蛋白質結構 (protein structure) ...................................................................... 1 1.1-1 蛋白質的三維結構 ...................................................................................... 1 1.2 蛋白質摺疊 (protein folding) .......................................................................... 7 1.3 寡聚蛋白質的摺疊與聚集 (oligomeric protein folding and association) ...................................................................................................................... 11 1.4 抗菌胜肽(antimicrobial peptides) ................................................................ 12 1.5 Mastoparan-B (MP-B) 介紹 ............................................................................ 13 1.6 研究目的 ................................................................................................................. 14 第二章 實驗原理................................................................................................... 15 2.1 固相胜肽合成法 (Solid-Phase Peptide Synthesis, SPPS) ................ 15 2.2 圓二色旋光光譜儀 (Circular Dichroism Spectrometer, CD) ......... 20 2.2-1 圓二色光譜儀原理 .................................................................................... 20 2.2-2 多胜肽的各種二級結構與其CD 光譜 ............................................ 22 2.2-3 計算α-helix 含量 ...................................................................................... 23 2.3 二維核磁共振 (Two dimensional nuclear magnetic resonance, 2D NMR) ................................................................................................................................. 24 2.3-1 Correlation Spectroscopy, COSY .......................................................... 28 2.3-2 Total Correlation Spectroscopy, TOCSY ........................................... 31 2.3-3 Nuclear Overhauser Effect Spectroscopy, NOESY ........................ 33 2.4 Diffusion Ordered Spectroscopy, DOSY .................................................... 35 2.4-1 擴散 (Diffusion) ......................................................................................... 35 2.4-2 脈衝磁場梯度核磁共振 (Pulsed field gradient NMR, PFG-NMR) .................................................................................................................. 35 2.4-3 自結合(Self-association) ................................................................... 36 2.5 分子動力學 (Molecular dynamics) ............................................................ 38 2.5-1 NMR 弛緩 (NMR Relaxation) .............................................................. 38 2.5-2 NMR 弛緩與動力學的關係.................................................................... 39 2.5-3 弛緩參數 (Relaxation Parameters) 和光譜密度函數 (spectral density function) 之間的關係 ......................................................... 40 2.5-4 無模型法則 (Model-free approach) ................................................... 42 第三章 實驗材料與方法..................................................................................... 46 3.1 實驗材料 ................................................................................................................. 46 3.2 實驗方法 ................................................................................................................. 51 3.2-1 固相胜肽合成 .............................................................................................. 52 3.2-2 胜肽純化與分子量鑑定 .......................................................................... 54 3.2-3 圓二色旋光光譜儀 .................................................................................... 56 3.2-4 抗菌活性 ........................................................................................................ 57 3.2-5 核磁共振實驗 .............................................................................................. 60 3.2-6 結構計算 ........................................................................................................ 65 3.2.7 無模型法則 (Model-free approach) 計算....................................... 69 3.2-8 黏度實驗 ........................................................................................................ 73 第四章 實驗結果................................................................................................... 74 4.1 圓二色旋光光譜 .................................................................................................. 74 4.2 抗菌活性 ................................................................................................................. 77 4.3 NMR 光譜的判定 ................................................................................................ 80 4.4 DOSY 光譜 ........................................................................................................... 126 4.5 結構計算 ............................................................................................................... 137 4.6 胜肽分子的動力學行為 ................................................................................ 143 第五章 討論 .......................................................................................................... 147 5.1 利用 CD 和NMR 研究胜肽的螺旋摺疊結構 .................................... 147 5.2 改變胜肽C 末端殘基對於其疏水性和疏水力矩的影響 ............... 152 5.2-1 疏水性 (H) .................................................................................................. 154 5.2-2 疏水力矩 (μH) .......................................................................................... 157 5.3 胜肽的自結合狀態對於螺旋摺疊的影響 ............................................. 159 5.4 胜肽的兩親性對於抗菌活性的重要性 .................................................. 161 5.5 胜肽的動力學行為........................................................................................... 162 第六章 結論 .......................................................................................................... 164 第七章 參考資料................................................................................................. 166 圖目錄 圖1-1:天然胺基酸的結構示意圖。 ..................................................................... 2 圖1-2:20 種天然胺基酸結構。 .............................................................................. 3 圖1-3:胺基酸利用胜肽鍵連結形成長鏈聚合物。 ...................................... 4 圖1-4:蛋白質的二級結構。 .................................................................................... 4 圖1-5:Ramachandran plot。 ..................................................................................... 5 圖1-6:蛋白質的三級結構。 .................................................................................... 5 圖1-7:蛋白質的四級結構。 .................................................................................... 6 圖1-8:蛋白質的摺疊機制。 .................................................................................... 9 圖1-9:漏斗能量表面。 ............................................................................................ 10 圖1-10:蛋白質大小和時間尺度的關係圖。 ................................................. 10 圖2-1:利用連接試劑將胜肽的C 端接到Resin 上。 ................................ 16 圖2-2:連接試劑。 ...................................................................................................... 16 圖2-3:胺基酸的N 端加上Fmoc 保護基,以防止其他反應發生。 . 17 圖2-4:胺基酸側鏈的保護基。 ............................................................................. 17 圖2-5:胺基酸的活化與耦合。 ............................................................................. 18 圖2-6:胺基酸去保護。 ............................................................................................ 18 圖2-7:切除Resin 和側鏈上的保護基。 .......................................................... 19 圖2-8:光通過偏光過濾器產生平面偏極光。 ............................................... 20 圖2-9:偏極光示意圖。 ............................................................................................ 21 圖2-10:多胜肽的五種不同二級結構之CD 光譜。 .................................. 23 圖2-11:二維核磁共振實驗的脈衝序列。 ...................................................... 24 圖2-12:施加一個90°x 脈衝。 ............................................................................... 24 圖2-13:線性遞增的t1 時間。 ............................................................................... 25 圖2-14:FID 經傅立葉轉換,從time domain 轉換成frequency domain。 .................................................................................................................................................. 27 圖2-15:COSY 的脈衝序列。 ................................................................................ 28 圖2-16:COSY 光譜的雙自旋系統示意圖。 .................................................. 30 圖2-17:TOCSY 的脈衝序列。 ............................................................................. 31 圖2-18:TOCSY 光譜的自旋系統示意圖。.................................................... 32 圖2-19:雙自旋系統 (I、S) 的偶極耦合。 ................................................... 33 圖2-20:NOESY 的脈衝序列。 ............................................................................. 33 圖2-21:兩個相鄰胺基酸之間質子的距離。 ................................................. 34 圖2-22:局部與總體的物理模型示意圖。 ...................................................... 44 圖3-1:實驗流程圖。 ................................................................................................. 51 圖3-2:固相胜肽合成流程圖。 ............................................................................. 53 圖3-3:純化後MP-B1-13 的HPLC 層析圖。 ................................................... 55 圖3-4:MP-B1-13 的MALDI-TOF MS 圖譜。 ................................................. 55 圖3-5:理想的劑量反應曲線。 ............................................................................. 59 圖4-1:MP-B 在溫度300 K 下,不同TFE 濃度的CD 光譜。 ............ 74 圖4-2:胜肽在30% TFE / 70% H2O 溶液中,溫度分別為283 K、298 K、300 K 和310 K 的CD 光譜疊圖。 ............................................................... 75 圖4-3:MP-B1-13 對E.coli 的劑量反應曲線。 ................................................ 77 圖4-4:MP-B 對E.coli 的劑量反應曲線。 ...................................................... 78 圖4-5:MP-BL15 對E.coli 的劑量反應曲線。 ................................................. 78 圖4-6:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的TOCSY 光譜。 .................................................................................................................................................. 81 圖4-7:MP-B 在30% TFE-d3 / 70% H2O,310 K 的TOCSY 光譜。 . 82 圖4-8:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的TOCSY 光譜。 .................................................................................................................................................. 83 圖4-9:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα HN 交叉峰區塊。 ...................................................................................................... 84 圖4-10:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-HN 交叉峰區塊。 ...................................................................................................... 85 圖4-11:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-HN 交叉峰區塊。 ...................................................................................................... 86 圖4-12:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hβ-HN 交叉峰區塊。 ............................................................................................... 87 圖4-13:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hβ-HN 交叉峰區塊。 ...................................................................................................... 88 圖4-14:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hβ-HN 交叉峰區塊。 ...................................................................................................... 89 圖4-15:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的HN-HN 交叉峰區塊。 ............................................................................................... 90 圖4-16:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 HN-HN 交叉峰區塊。 ..................................................................................................... 91 圖4-17:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 HN-HN 交叉峰區塊。 ..................................................................................................... 92 圖4-18:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hα-HN 交叉峰區塊。 ............................................................................................... 94 圖4-19:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-HN 交叉峰區塊。 ...................................................................................................... 95 圖4-20:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-HN 交叉峰區塊。 ...................................................................................................... 96 圖4-21:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hβ-HN 交叉峰區塊。 ............................................................................................... 97 圖4-22:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hβ-HN 交叉峰區塊。 ...................................................................................................... 98 圖4-23:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hβ-HN 交叉峰區塊。 ...................................................................................................... 99 圖4-24:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hγ-HN 交叉峰區塊。.............................................................................................. 100 圖4-25:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hγ-HN 交叉峰區塊。 .................................................................................................... 101 圖4-26:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hγ-HN 交叉峰區塊。 .................................................................................................... 102 圖4-27:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hα-Hβ 交叉峰區塊。 .............................................................................................. 103 圖4-28:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-Hβ 交叉峰區塊。 .................................................................................................... 104 圖4-29:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-Hβ 交叉峰區塊。 .................................................................................................... 105 圖4-30:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 的Hα-Hγ 交叉峰區塊。顯示dαγ(i, i+3)的NOE 訊號。 .............................. 106 圖4-31:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-Hγ 交叉峰區塊。 .................................................................................................... 107 圖4-32:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜的 Hα-Hγ 交叉峰區塊。 .................................................................................................... 108 圖4-33:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 芳香環側鏈區域。 ....................................................................................................... 109 圖4-34:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜芳香 環側鏈區域。 ................................................................................................................. 110 圖4-35:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜芳 香環側鏈區域。 ............................................................................................................ 111 圖4-36:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜 尾端胺基區域。 ............................................................................................................ 112 圖4-37:MP-B 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜尾端 胺基區域。 ....................................................................................................................... 113 圖4-38:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的NOESY 光譜尾 端胺基區域。 ................................................................................................................. 114 圖4-39:二次化學位移 (secondary chemical shift)。 ............................... 119 圖4-40:MP-B1-13 在30% TFE-d3 / 70% H2O,溫度310 K 時的NOE 訊號。 ................................................................................................................................. 120 圖4-41:MP-B 在30% TFE-d3 / 70% H2O,溫度310 K時的NOE 訊號。 ................................................................................................................................................ 121 圖4-42:MP-BL15 在30% TFE-d3 / 70% H2O,溫度310 K 時的NOE 訊 號。 ...................................................................................................................................... 121 圖4-43:MP-B1-13 在30% TFE-d3 / 70% H2O,溫度310 K 的1H-13C HSQC 光譜。 .................................................................................................................. 122 圖4-44:MP-B 在30% TFE-d3 / 70% H2O,溫度310 K 的1H-13C HSQC 光譜。 ................................................................................................................................. 123 圖4-45:MP-BL15 在30% TFE-d3 / 70% H2O,溫度310K的1H-13C HSQC 光譜。 ................................................................................................................................. 124 圖4-46:MP-B1-13 在30% TFE-d3 / 70% H2O,八個不同溫度下的DOSY 光譜疊圖。 ....................................................................................................................... 127 圖4-47:MP-B 在30% TFE-d3 / 70% H2O,八個不同溫度下的DOSY 光譜疊圖。 ....................................................................................................................... 128 圖4-48:MP-BL15 在30% TFE-d3 / 70% H2O,八個不同溫度下的DOSY 光譜疊圖。 ....................................................................................................................... 129 圖4-49:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 下的DOSY 光譜 圖。 ...................................................................................................................................... 130 圖4-50:MP-B 在30% TFE-d3 / 70% H2O,310 K 下的DOSY 光譜圖。 ................................................................................................................................................ 131 圖4-51:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 下的DOSY 光譜 圖。 ...................................................................................................................................... 132 圖4-52:MP-B1-13、MP-B 和MP-BL15 在30% TFE-d3 / 70% H2O 溶液 中,不同溫度下的擴散係數。 .............................................................................. 136 圖4-53:MP-B1-13、MP-B 和MP-BL15 在30% TFE-d3 / 70% H2O 溶液 中,不同溫度下的自結合狀態。 .................................................................. 136 圖4-54:MP-B1-13 在100 個結構之中四種主要的螺旋摺疊結構,其骨 幹的疊圖。 ....................................................................................................................... 138 圖4-55:MP-B 在100 個結構之中四種主要的螺旋摺疊結構,其骨幹 的疊圖。 ............................................................................................................................ 139 圖4-56:MP-BL15 在100 個結構之中四種主要的螺旋摺疊結構,其骨 幹的疊圖。 ....................................................................................................................... 140 圖4-57:20 個最低能量結構的Ramachandran plot 圖。 ........................ 141 圖5-1:MP-B1-13、MP-B 與MP-BL15 的隨著溫度變化的α-helix 含量 (%)。 ................................................................................................................................... 147 圖5-2:C 末端胺基跟其他殘基之間的氫鍵長度。 ................................... 150 圖5-3:MP-B-COO-沒有尾端胺基與其他殘基之間的氫鍵。 .............. 151 圖5-4:滯留係數與HPLC 滯留時間的關係圖。 ....................................... 156 圖5-5:HPLC 滯留時間與疏水性的關係圖。 .............................................. 156 圖5-6:胜肽的兩親性示意圖。 ........................................................................... 158 圖5-7:MP-BL15 每個殘基的次序參數 (S2)。 .............................................. 163 圖5-8:MP-BL15 的假想的雙體模型。 .............................................................. 163 表目錄 表1-1:重要的類澱粉蛋白疾病及相關的蛋白質。 .............................. 7 表2-1:CD 光譜中多胜肽的五種不同二級結構之特定吸收波長。 . 22 表2-2:弛緩參數與光譜密度函數之間的關係。 ................................ 41 表2-3:Model 1 ~ 5 所對應的的動力學參數。 ................................... 44 表3-1:合成時所設定的活化、偶合、去保護時間。 ........................ 52 表3-2:純化樣品時HPLC 的梯度設定。 .......................................... 54 表3-3:圓二色旋光光譜儀的實驗參數。 ............................................ 56 表3-4:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的參數設定。 .. 61 表3-5:MP-B 在30% TFE-d3 / 70% H2O,310 K 的參數設定。 ...... 61 表3-6:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的參數設定。 ... 62 表3-7:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 弛緩實驗的參數 設定。 ....................................................................................................... 62 表3-8:MP-B在30% TFE-d3 / 70% H2O,310 K 弛緩實驗的參數設定。 ................................................................................................................... 63 表3-9:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 弛緩實驗的參數設 定。 ........................................................................................................... 63 表3-10:T1 和T2 的延遲時間。 ............................................................ 64 表3-11:mfinput 變數。 ........................................................................ 69 表3-12:實驗溫度下所對照的純水黏度值 (cP)。 ............................. 73 表4-1:MP-B1-13、MP-B 和MP-BL15 不同溫度下的α-helix 含量。 . 76 表4-2:MP-B1-13、MP-B、MP-BL15 和MP-B-COO-的半抑制濃度 (IC50)。 ..................................................................................................... 79 表4-3:MPB1-13 的1H 化學位移表。濃度5 mM,在30% TFE-d3 / 70% H2O 溶液中,pH 3.70,溫度310 K。 ................................................ 115 表4-4:MPB 的1H 化學位移表。濃度3.7 mM,在30% TFE-d3 / 70% H2O 溶液中,pH 4.08,溫度310 K。 ................................................ 116 表4-5:MPBL15,濃4 mM,在30% TFE-d3 / 70% H2O 溶液中,pH 3.75, 溫度310 K 時的1H 化學位移表。 ...................................................... 117 表4-6:20 種常見的胺基酸在無序纏繞結構下,Hα 的化學位移範圍。 ................................................................................................................. 118 表4-7:MP-B1-13、MP-B 和MP-BL15 的13Cα 化學位移表。在30% TFE-d3 / 70% H2O 溶液中,溫度310 K。 ....................................................... 125 表4-8:MP-B1-13 在30% TFE-d3 / 70% H2O 的溶液中,各溫度下的擴 散系數 (D)、溶液黏度 (η) 及分子量。 ............................................ 133 表4-9:MP-B 在30% TFE-d3 / 70% H2O 的溶液中,各溫度下的擴散 系數 (D)、溶液黏度 (η) 及分子量。 ................................................ 134 表4-10:MP-BL15 在30% TFE-d3 / 70% H2O 的溶液中,各溫度下的擴 散系數 (D)、溶液黏度 (η) 及分子量。 ............................................ 135 表4-11:MP-B1-13、MP-B 和MP-BL15 和MP-B-COO-經XPLOR 計算 後的100 個結構統計表。 .................................................................... 142 表4-12:MP-B1-13、MP-B 和MP-BL15 的20 個最低能量結構的RMSD 值 (A )。 ................................................................................................. 142 表4-13:MP-B1-13 在30% TFE-d3 / 70% H2O,310 K 的弛緩參數R1、 R2、NOE 和次序參數S2。 ................................................................... 144 表4-14:MP-B 在30% TFE-d3 / 70% H2O,310 K 的弛緩參數R1、R2、 NOE 和次序參數S2。 ........................................................................... 145 表4-15:MP-BL15 在30% TFE-d3 / 70% H2O,310 K 的弛緩參數R1、 R2、NOE 和次序參數S2。 ................................................................... 146 表5-1:比較MP-B1-13 和MP-B 在283 K 和310 K 的dNN(i, i+1) NOE 訊號。 ..................................................................................................... 148 表5-2:Eisenberg plot 的疏水尺度。此疏水尺度是統合了其他文獻的 結果得到的值。 .................................................................................. 153 表5-3:Fauchere and Dliska 的疏水尺度。 ........................................ 153 表5-4:Whole-Residue 疏水尺度 。從水到POPC 界面的轉移自由能 (free energies of transfer, ΔG (kcal/mol))。 .......................................... 154 表5-5:胺基酸的滯留係數。 .............................................................. 155 |
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