本文關(guān)鍵詞：天然糖鏈的提取和糖肽的合成　出處：《南昌大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 糖苷內(nèi)切酶法 寡糖鏈 糖鏈VA唑啉 多肽受體 酶催化糖基化

【摘要】：蛋白的糖基化是蛋白質(zhì)翻譯后修飾的一種主要的形式。由于糖蛋白和糖肽的糖鏈的不均一性,導(dǎo)致其功能上的顯著差異。因此,合成均一的糖蛋白和糖肽對(duì)于糖鏈結(jié)構(gòu)和功能的研究是必不可少的。本論文探索了從核糖核酸酶B(RB)、唾液酸糖肽(SGP)和雞卵清白蛋白(OVA)三種天然來(lái)源糖蛋白/糖肽來(lái)制備糖鏈底物的方法,并成功制備了高甘露糖型和復(fù)合型兩種N-糖鏈類型的VA唑啉底物。我們選擇了10條多肽序列并合成了帶乙酰氨基葡萄糖(GlcNAc)的多肽受體,利用了酶催化方法制備了14條高甘露糖型和復(fù)合糖型的糖肽。完善了酶催化糖肽合成方法;建立了從RB中提取高甘露糖型糖鏈及制備相應(yīng)VA唑啉的半合成方法;探索了從OVA提取糖鏈的工藝條件和分離條件;制備了可用于糖蛋白質(zhì)組學(xué)分析的標(biāo)準(zhǔn)糖肽。論文第一章概述了糖肽的合成方法、糖苷內(nèi)切酶法合成糖肽策略的研究以及糖肽化合物的應(yīng)用。第二章主要敘述了通過(guò)半合成法合成糖鏈供體底物。利用全合成法合成步驟比較繁瑣不易純化,而且很難得到完整的寡糖鏈供體。本文通過(guò)從天然蛋白中分離提取寡糖鏈,并通過(guò)一鍋法合成VA唑啉底物,步驟很簡(jiǎn)短,在分離方面可以減少產(chǎn)物的損失,可以高效地得到糖鏈給體底物。論文第三章通過(guò)Fmoc固相合成法合成了一系列含有GlcNAc的Fmoc保護(hù)的天冬酰胺的多肽受體底物。利用Fmoc固相合成法可以簡(jiǎn)便、快速和高效地得到多肽。第四章通過(guò)酶催化反應(yīng)將合成的糖鏈供體底物轉(zhuǎn)移到含有GlcNAc的Fmoc保護(hù)的天冬酰胺的多肽受體上,高效地合成了均一結(jié)構(gòu)的糖肽。在本章中我們探索了利用天然糖蛋白中提取的并合成新的寡糖鏈VA唑啉作為糖基供體合成均一糖肽的方法。
[Abstract]:Protein glycosylation is one of the main forms of post-translational modification. Due to heterogeneity of the sugar chain glycoproteins and glycopeptides, lead to significant differences of its function. Therefore, the research on the structure and function of sugar chain synthesis homogeneous glycoproteins and glycopeptides is essential. This thesis explores from ribonuclease B (RB), sialoglycopeptide (SGP) and ovalbumin (OVA) three natural sources of glycoproteins / glycopeptides method for preparing carbohydrate substrates, and prepared VA quinazoline substrate high mannose type and compound type two N- sugar chain type. We chose 10 polypeptide sequences with the synthesis of acetyl glucosamine (GlcNAc) polypeptide receptor, using enzymatic methods for 14 high mannose type and complex type sugar glycopeptides obtained. Improved enzyme catalyzed glycopeptide synthesis method; a preparation phase high mannose type glycans and extracted from RB Semi synthesis method of VA quinazoline; to explore the extraction process of sugar chain from OVA and separation conditions; the preparation can be used for standard glycopeptide sugar proteomics analysis. The first chapter summarizes the synthesis of glycopeptides of endoglycosidase synthesis and application of compound glycopeptide glycopeptide strategy. The second chapter described by semi synthesis of sugar chain donor substrate. Using synthesis steps are more complex and difficult purification, it is difficult to complete the oligosaccharide chain donor. Through extraction and separation of oligosaccharides from natural protein, and through one pot synthesis of oxazoline VA substrate step is very short, the separation can be reduced the product can effectively obtain the loss of sugar chain donor substrates. In the third chapter, the synthesis of a series of Fmoc GlcNAc containing asparagine polypeptide receptor substrate by Fmoc solid-phase synthesis method using. Fmoc solid phase synthesis method can be simple, fast and efficient peptidemaintained. In the fourth chapter, by enzyme catalytic reaction to synthesis of sugar chain asparagine donor substrate transfer to polypeptide receptor Fmoc protection containing GlcNAc on the efficient synthesis of uniform structure of the glycopeptide. In this chapter we explore the method of extracting natural glycoprotein in use and the synthesis of new VA oligosaccharide chain oxazoline as glycosyl donor glycopeptide synthesis of uniform.

【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：O629

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相關(guān)期刊論文 前2條

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本文編號(hào)：1360520