合成具有生物活性的醣蛋白質及醣類改質

Synthesis of biological active glycoprotein and polysaccharide modification

陳正晞(Cheng-Hsi Chen)

指導教授 ： 華國媛

國立臺北科技大學/工程學院/有機高分子研究所/碩士(2006年)

摘要

Polysaccharide有許多生物功能性，對於細胞來說是一個非常重要的巨分子，例如細胞間的訊息傳遞、吸收水分以維持組織間的水分。 本實驗是設計有功能性的新穎醣蛋白質，首先利用一些polysaccharides進行氧化反應，由於氧化過後的polysaccharide之 hydroxyl group氧化成具有反應性的 aldehyde group，則可以和4(4-N-maleimidophenyl)butyric acid hydrazide.HCl.1/2 dioxane (MPBH)。此種linker上的hydrazide group 形成共價鍵結，之後我們再讓BSA之 thiol group和MPBH上的maleimide group形成共價鍵結，合成出來的compounds 為polysaccharides conjugated BSA。 因為BSA為一種攜帶蛋白質，可以利用endocytosis 的作用將polysaccharide 送入細胞內，所以polysaccharides conjugated BSA可以作為免疫細胞的刺激劑來刺激免疫反應。我們選用mannan，pectin此兩種polysaccharides來進行conjugate BSA的反應，之後利用lectin blot以及periodic acid Schiff (PAS) staining來驗證polysaccharides 是否有無conjugated BSA。之後我們將測試多醣體之生物功能。
除此之外，我們進行多醣體的功能性，利用化學合成改質其C-6位置的官能基，讓原本的hydroxyl group 改質成 xanthate group和xanthate acetylation group。之後化合物結構分析鑑定使用GC-MS, IR 1HNMR 和13CNMR來測定其改質過後的官能基。我們將進行polysaccharide xanthate和polysaccharide xanthate acetylation之生物活性測試。

關鍵字

BSA ； 4(4-N-maleimidophenyl)butyric acid hydrazide.HCl.1/2 dioxane (MPBH) ； Lectin ； Polysaccharide ； Xanthate

並列摘要

Polysaccharides are important macromolecules with many cellular functions, such as transmitting signals between cells and absorbing waters to maintain moist of tissues. Our project was designed to elicit the functions of neoglycoconjugates. First, polysaccharides were oxidized into aldehyde group. The treatment allowed the sequential reaction, to form covalent bond with 4(4-N-maleimidophenyl) butyric acid hydrazide.HCl.1/2 dioxane. The glycan then was cross linked to BSA, as a protein carrier with maleimide group of 4(4-N-maleimidophenyl) butyric acid hydrazide.HCl.1/2 dioxane. The synthesized compounds were glycoconjugated-BSA. To detect the polysaccharide conjugates, we used lectin blot and periodic acid Schiff (PAS) staining. We then tested the biological functions of polysaccharides. This allowed glycoconjugtates to be taken up by a cell through endocytosis. Glycoconjugate-BSA was able to stimulate immune cells as immuno-modulator.
To test the functions of polysaccharides,modification on C-6 hydroxyl group into xanthate and xanthate acetylation. To detect the modified functional group, GC-MS, IR and 1HNMR, 13CNMR were used. We will test the biological functions of these polysaccharides in the future.

並列關鍵字

BSA ； 4(4-N-maleimidophenyl)butyric acid hydrazide.HCl.1/2 dioxane (MPBH) ； Lectin ； Polysaccharide ； Xanthate

參考文獻

2. Rollag, H. and M. Degre, Effect of recombinant interferon-gamma on protein content, phagocytic, and cytotoxic activity of mouse peritoneal macrophages. J Interferon Res, 1988. 8(2): p. 169-78.

3. Almond, A. and J.K. Sheehan, Predicting the molecular shape of polysaccharides from dynamic interactions with water. Glycobiology, 2003. 13(4): p. 255-64.

4. San Blas, G. and W.L. Cunningham, Structure of cell wall and exocellular mannans from the yeast Hansenula holstii. I. Mannans produced in phosphate-containing medium. Biochim Biophys Acta, 1974. 354(2): p. 233-46.

5. Chakraborty, P., A.N. Bhaduri, and P.K. Das, Neoglycoproteins as carriers for receptor-mediated drug targeting in the treatment of experimental visceral leishmaniasis. J Protozool, 1990. 37(5): p. 358-64.

6. Hille, A., A. Waheed, and K. von Figura, Assembly of the ligand-binding conformation of Mr 46,000 mannose 6-phosphate-specific receptor takes place before reaching the Golgi complex. J Cell Biol, 1990. 110(4): p. 963-72.