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系統識別號 U0002-2507201414453100
中文論文名稱 C末端殘基如何改變兩親性螺旋摺疊:CD和NMR的研究
英文論文名稱 How C-terminal residue alters the helix folding of an amphipathic antimicrobial peptide: CD and NMR study
校院名稱 淡江大學
系所名稱(中) 化學學系碩士班
系所名稱(英) Department of Chemistry
學年度 102
學期 2
出版年 103
研究生中文姓名 曾雅琳
研究生英文姓名 Ya-Lin Tseng
學號 601160061
學位類別 碩士
語文別 中文
口試日期 2014-07-10
論文頁數 170頁
口試委員 指導教授-李長欣
委員-李長欣
委員-鄧金培
委員-錢嘉琳
中文關鍵字 C末端  兩親性  摺疊  CD  NMR 
英文關鍵字 Folding  antimicrobial peptide  C-terminal  CD  NMR 
學科別分類 學科別自然科學化學
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.
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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