本文選題：唾液 + 血清��； 參考：《西北大學(xué)》2013年碩士論文

【摘要】：流行性感冒(Influenza)是一種由流感病毒侵染機(jī)體而引發(fā)的急性呼吸道傳染病。已有研究證實(shí),甲型流感病毒血凝素(HA)分子與流感病毒宿主細(xì)胞表面SAα2-3Gal或SAα2-6Gal糖鏈末端受體的結(jié)合,是流感病毒侵染機(jī)體的開始。不同流感病毒宿主細(xì)胞表面上的糖鏈?zhǔn)荏w結(jié)構(gòu)是不同的,禽流感病毒血凝素(HA)主要識別和結(jié)合末端為SAα2-3Gal的糖鏈?zhǔn)荏w,而人流感病毒血凝素(HA)主要識別和結(jié)合末端為SAα2-6Gal的糖鏈?zhǔn)荏w。研究報道指出SAα2-6Gal糖鏈末端結(jié)構(gòu)主要存在于人上呼吸道細(xì)胞表面,在人下呼吸道細(xì)胞表面僅有少量SAα2-3Gal糖鏈末端結(jié)構(gòu)的存在。這些發(fā)現(xiàn)為抗流感病毒的相關(guān)研究提供了全新的思路,提示SAα2-3Gal和SAα2-6Gal糖鏈結(jié)構(gòu)可以作為分離純化血凝素,以及能夠與血凝素競爭性結(jié)合該糖鏈?zhǔn)荏w糖結(jié)合蛋白的工具。本研究以此為思路,利用SAα2-3Gal和SAα2-6Gal糖鏈結(jié)構(gòu)分別從健康人唾液和血清中分離及鑒定抗流感病毒保護(hù)性蛋白,并結(jié)合生物信息學(xué)研究手段對這些蛋白質(zhì)的相關(guān)信息進(jìn)行深入分析,尋找這些蛋白質(zhì)與糖鏈SAα2-3Gal和SAα2-6Gal結(jié)合的結(jié)構(gòu)域。 主要參考本實(shí)驗(yàn)室已經(jīng)建立的羥基化磁性微粒分離純化糖結(jié)合蛋白的方法,利用SAα2-3Gal和SAα2-6Gal糖鏈修飾的功能化磁性微粒,從健康人唾液和血清中分別分離純化糖鏈SAα2-3Gal和SAα2-6Gal結(jié)合蛋白(抗流感病毒保護(hù)性),并對這些蛋白質(zhì)進(jìn)行質(zhì)譜鑒定；通過質(zhì)譜數(shù)據(jù)分析唾液和血清中SAα2-3Gal和SAα2-6Gal結(jié)合蛋白的異同,為進(jìn)一步研究人體內(nèi)抗流感病毒保護(hù)性蛋白質(zhì)在預(yù)防流感傳播中的作用提供理論依據(jù)；最后利用motif-x基序分析模型,預(yù)測了特異性結(jié)合SAα2-3Gal和SAα2-6Gal糖鏈蛋白質(zhì)的糖結(jié)合域。 對獲得的質(zhì)譜數(shù)據(jù)進(jìn)行整理、篩選、統(tǒng)計和分析,得到以下實(shí)驗(yàn)結(jié)果：1)從健康人唾液中共鑒定到116個SAα2-3Gal和SAα2-6Gal糖結(jié)合蛋白,其中94個SAα2-3Gal糖結(jié)合蛋白,83個SAα2-6Gal糖結(jié)合蛋白,與這兩種糖鏈都結(jié)合的蛋白質(zhì)有61個,與SAα2-3Gal糖鏈特異性結(jié)合蛋白質(zhì)有33個,與SAα2-6Gal糖鏈特異性結(jié)合蛋白質(zhì)有22個；兩組唾液糖結(jié)合蛋白質(zhì)的相對分子質(zhì)量和等電點(diǎn)分布較為類似；由emPAI分析結(jié)果可知,Ratio Value(即mol%(SA2-6Gal):mol%(SAα2-3Gal)大于1.5的蛋白質(zhì)有35個,小于0.66的蛋白質(zhì)有9個；2)從健康人血清中共鑒定到144個SAα2-3Gal和SAα2-6Gal糖結(jié)合蛋白,其中117個SAα2-3Gal糖結(jié)合蛋白,115個SAα2-6Gal糖結(jié)合蛋白,與這兩種糖鏈都結(jié)合的蛋白質(zhì)有88個,與SAα2-3Gal糖鏈特異性結(jié)合蛋白質(zhì)有29個,與SAa2-6Gal糖鏈特異性結(jié)合蛋白質(zhì)有27個；兩組血清糖結(jié)合蛋白的相對分子質(zhì)量和等電點(diǎn)分布較為類似；由emPAI分析結(jié)果可知,Ratio Value(即mol%(SAα2-6Gal): mol%(SAα2-3Gal))大于1.5的蛋白質(zhì)有52個,小于0.66的蛋白質(zhì)有23個；3)統(tǒng)計分析可知,唾液和血清中與這兩種糖鏈同時結(jié)合的蛋白質(zhì)有13種,與SAα2-6Gal糖鏈同時結(jié)合的蛋白質(zhì)僅有2種,分別為免疫球蛋白G-λ2鏈C區(qū)和免疫球蛋白G-κ鏈Ⅴ-Ⅳ區(qū)：與SAα2-3Gal糖鏈結(jié)合的蛋白質(zhì)未出現(xiàn)重疊。根據(jù)motif-x基序分析可知,SAα2-3Gal糖鏈特異性結(jié)合蛋白質(zhì)的糖結(jié)合潛在特征性糖結(jié)合域有44種,而SAα2-6Gal糖鏈特異性結(jié)合蛋白的糖結(jié)合潛在特征性糖結(jié)合域只有1種。
[Abstract]:Influenza virus (Influenza) is an acute respiratory infection caused by influenza virus infection. It has been proved that the binding of HA molecules with SA alpha 2-3Gal or SA alpha 2-6Gal sugar chain terminal receptor on the surface of the host cell of influenza virus is the beginning of influenza virus infection. The structure of the sugar chain receptor on the surface of the main cell is different, and avian influenza virus hemagglutinin (HA) mainly identifies and combines the sugar chain receptor at the end of SA alpha 2-3Gal, while human influenza virus hemagglutinin (HA) mainly identifies and combines the end of the glucose chain receptor with SA alpha 2-6Gal. The study shows that the terminal structure of the SA alpha 2-6Gal chain is mainly in the human upper respiratory tract. On the surface of the cell, only a small amount of SA alpha 2-3Gal sugar chain terminal structure exists on the surface of the human lower respiratory tract. These findings provide a new idea for the related research of anti influenza virus, suggesting that SA alpha 2-3Gal and SA alpha 2-6Gal sugar chain structure can be used to separate and purify hemagglutinin, and to combine with hemagglutinin to combine the sugar chain receptor with the hemagglutinin. In this study, we use SA alpha 2-3Gal and SA alpha 2-6Gal sugar chain structure to separate and identify anti influenza virus protective proteins from healthy human saliva and serum, and analyze the related information of these proteins in depth with bioinformatics research methods, and find these proteins and sugar chain SA alpha 2-3Gal and S. The domain of A alpha 2-6Gal binding.
We mainly refer to the method of separating and purifying the sugar binding protein by the hydroxy magnetic particles established in our laboratory, and using the functional magnetic particles modified by SA alpha 2-3Gal and SA alpha 2-6Gal sugar chain to separate and purify the sugar chain SA alpha 2-3Gal and SA alpha 2-6Gal in combination with the egg white (anti influenza virus protection) from the healthy human saliva and serum. Mass spectrometric identification and analysis of the similarities and differences of SA alpha 2-3Gal and SA alpha 2-6Gal binding protein in saliva and serum by mass spectrometry, provide a theoretical basis for further research on the role of anti influenza virus protective proteins in the prevention of influenza transmission. Finally, the motif-x based sequence analysis model is used to predict the specific binding of SA alpha 2-3Gal. The sugar binding domain of SA alpha 2-6Gal sugar chain protein.
The obtained mass spectrometry data were collated, screened, statistics and analyzed. 1) 116 SA alpha 2-3Gal and SA alpha 2-6Gal binding proteins were identified from healthy human saliva, of which 94 SA alpha 2-3Gal sugar binding proteins, 83 SA alpha 2-6Gal sugar binding proteins, and 61 proteins combined with these two sugars, and SA alpha 2-3Gal sugar. There are 33 chain specific protein binding proteins and 22 proteins associated with SA alpha 2-6Gal sugar chain specific binding proteins; the relative molecular mass and isoelectric point distribution of the two groups of saliva sugar binding proteins are similar; the results of emPAI analysis show that there are 35 proteins of Ratio Value (mol% (SA2-6Gal): mol% (SA alpha 2-3Gal) greater than 1.5, less than 0.66 of protein 9; 2) 144 SA alpha 2-3Gal and SA alpha 2-6Gal binding proteins were identified from healthy human serum, of which 117 SA alpha 2-3Gal sugar binding proteins, 115 SA alpha 2-6Gal sugar binding proteins, 88 proteins combined with these two sugar chains, 29 with SA alpha 2-3Gal sugar chain specific binding egg white, and specific binding protein of SAa2-6Gal sugar chain. The relative molecular mass and isoelectric point distribution of serum sugar binding proteins in the two groups were similar, and the results of emPAI analysis showed that Ratio Value (mol% (SA alpha 2-6Gal): mol% (SA a 2-3Gal)) had 52 proteins greater than 1.5, 23 protein less than 0.66; 3) statistical analysis showed that saliva and serum were associated with the two sugar chains. There are 13 kinds of proteins combined with the protein of SA alpha 2-6Gal sugar chain. There are only 2 proteins, which are the immunoglobulin G- lambda 2 chain C region and the immunoglobulin G- kappa chain V - IV region: the protein binding to the SA alpha 2-3Gal chain is not overlapped. According to the motif-x motif analysis, the sugar chain specific binding protein of SA alpha 2-3Gal is potential for sugar binding. There are 44 kinds of characteristic sugar binding domains, while the sugar binding potential characteristic sugar binding domain of SA alpha 2-6Gal sugar chain binding protein is only 1.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：Q51;R511.7

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