利用多維核磁共振技術對人類Interleukin-1α水溶液結構之探討

Study on the structure of human Interleukin-1α by using multi-dimension NMR technology

張軒國(Hsuan-Kuo Chang)

指導教授 ： 余靖

國立清華大學/理學院/化學系/碩士(2009年)

摘要

IL-1α(介白素,Interleukin-1α)為一具有多樣生物活性的訊息傳遞分子，其經由非典型路徑分泌出細胞後，會與細胞表面的受器作用並引發訊息傳遞。如引起發燒、刺激肝臟製造急性蛋白、增大淋巴細胞的回應、誘發關節退化、增加骨髓細胞數目等等；同時亦會誘導數種腦丘下部和腦垂體多肽的釋放及胰島素的轉錄。且IL-1α也是造成類風濕性關節炎與退化性關節炎的主因之一，其在人體中扮演了重要的角色。
本篇論文中，主要利用多維核磁共振技術解出IL-1α在水溶液中的結構。首先使用各種三維核磁共振實驗如HNCA、HN(CO)CA、HNCO來得到IL-1α蛋白分子的骨架資訊；CBCA(CO)NH、CBCANH、HBHA(CO)NH、HCCH-TOCSY來得到IL-1α蛋白分子的支鏈資訊，以這些結果判定95% 胺基酸中的1H、15N和13C的化學位移。進行結構計算時，先由15N & 13C-edited NOESY來得到IL-1α蛋白分子內所有NOE的資訊，並且配合雙面角、氫鍵、CSI等限制條件，接著使用Aria/CNS軟體來解出IL-1α蛋白分子在水溶液中的結構，以作為研究IL-1α與S100A13蛋白複合結構之基礎，此複合結構可幫助我們更深入的了解IL-1α的non-classical pathway。

關鍵字

核磁共振 ； 水溶液結構

並列摘要

Interleukin-1α is one of the members of signaling transduction molecules which serves diverse biological functions. It is secreted by the cell through the non-classical pathway and interacts with the receptor on the cell surface to trigger the signal transduction cascade. It could induce fever, stimulate hepatic acute-phase protein production, augment lymphocyte responses, induce degenerative changes in joints and increase the numbers of bone marrow cells etc. It is also able to induce the release of several kinds of hypothalamic and pituitary peptides and to enhance the transcription of insulin. IL-1α thus plays an important role in human body and is the primary element to cause rheumatic arthritis and degeneration arthritis.
In the thesis, the solution structure of IL-1α was solved using multi-dimensional nuclear magnetic resonance technology. First, the backbone information of the IL-1α was determined from a variety of 3D NMR experiments such as HNCA, HN(CO)CA, and HNCO, followed by other experiments such as CBCA(CO)NH, CBCANH, HBHA(CO)NH, and HCCH-TOCSY to determine the side-chain constraints of IL-1α. The 1H, 15N, 13C chemical shifts of 95% amino acid of IL-1α were successfully assigned.
The solution structure of IL-1α was calculated by Aria/CNS software program using the NOE constraints obtained from 15N & 13C-edited NOESY experiments. The other constraints such as dihedral angle, hydrogen bond, chemical shift index were also used for structure calculation etc. The 3D solution structure of IL-1α determined in the present study can help us to understand the non-classical pathway of IL-1α.

並列關鍵字

Interleukin-1α ； structure ； NMR

參考文獻

(1)Dinarello, C. A., (1988) Biology of interleukin 1. FASB J. 2：108-115

(2)Adrew J. P.,Steven E. H. (2009) Cytokine and cytokine receptor gene polymorphisms and their functionality. Cytokine & Growth Factor Reviews 20：43-59

(3)Furutani, Y., Notake, M., et al., (1986) Complete nucleotide sequence of the gene for human interleukin 1 alpha . Nucleic Acids Research 14：3167-79 ,

(4)Dinarello, C. A., (1994) The interleuldn-1 family: 10 years of discovery. FASB J. 8：1314-1325

(5)Mandrup-poulsen, T., Bendtzen, K., et al., (1986) Affinity-purified human interleukin-1 is cytotoxic to isolated islets of Langerhans. Diabetologia 29：63-67