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Please use this identifier to cite or link to this item:
https://tkuir.lib.tku.edu.tw/dspace/handle/987654321/32841
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| Title: | 利用CD和NMR研究合成的神經胜肽片段hNPY【21-31】之結構 |
| Other Titles: | Conformational studies of synthesized neuropeptide fragment hNPY[21-31] by CD and NMR. |
| Authors: | 林嘉豪;Lin, Chia-hao |
| Contributors: | 淡江大學化學學系碩士班
李長欣;Lee, Chang-shin |
| Keywords: | 二維核磁共振;圓二色光譜儀;神經胜肽;固相胜肽合成法;2D NMR;CD;NPY;PP;SPPS |
| Date: | 2008 |
| Issue Date: | 2010-01-11 02:50:18 (UTC+8) |
| Abstract: | 人類神經胜肽Y(Human Neuropeptide Y, hNPY)在水溶液中具有α-螺旋結構，且低濃度下結構為單體，高濃度時則為雙體。其結構有特定作用機制:單體下先與膜微脂粒(membrane micelle)結合後進一步與GPCRs(G protein-coupled receptors)作用。
固相胜肽合成法(Solid Phase Peptide Synthesis)合成hNPY片段序列(hNPY[21-31])，經由高效能液相層析儀純化，質譜儀確認分子量，再用圓二色光譜(Circular Dichroism)觀察不同比例TFE溶液下，二級結構的變化，選擇100%水溶液與50%TFE兩溶液做結構測量與討論。用二維核磁共振(two-dimensional NMR)的光譜: COSY、TOCSY、ROESY、NOESY、[1H, 13C]-HSQC及光譜判定(Assignment)和光譜循序判定(Sequential assignment)，各別判定兩溶液之11個殘基質子與13碳的化學位移。經由NOE限制條件分別計算兩溶液之3D結構，100%水溶液呈現鬆散不規則結構，50%TFE溶液則在24Leu~28Asn出現α-螺旋結構。
配合CD、CSI(Chemical Shift Index)、NMR、結構計算，討論兩比例溶液之結構差異，並與NPY結構比較。
Human Neuropeptide Y(hNPY) has a well-defined α-helical structure in solution, and is monomer at low concentration, dimer at high concentration. It has specific binding mechanism: First, the monomer structure binds with membrane micelle, and further interacts with G protein-coulpled receptors(GPCRs).
We synthesize neuropeptide fragment hNPY[21-31] by solid phase peptide synthesis, purified by RP-HPLC and made sure the molecular weight by Mass. The conformation and dynamics of hNPY[21-31] in difference solvent condition is studied by CD and 2D NMR experiment.
2D NMR experiments of COSY, TOCSY, ROESY, NOESY, and [1H, 13C]-HSQC were acquired. With NOE restrained structural calculation, the major structure of hNPY[21-31] in 100% H2O is random coil and form regular α-helical structure between 24Leu and 28Asn in 50%TFE/50% H2O.
Combination of CD, NMR, and XPLOR molecular calculation, we can investigate the conformational difference between 100%H2O and 50%TFE /50%H2O, and compare with native NPY. |
| Appears in Collections: | [化學學系暨研究所] 學位論文
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