本文選題：多肽 + 肝素�。� 參考：《吉林大學(xué)》2011年博士論文

【摘要】：人乳頭瘤病毒(HPV)是一類嗜上皮性無囊膜雙鏈DNA病毒,迄今已在人體皮膚及黏膜組織中分離出100多種不同型別。病毒的生命周期以感染細(xì)胞為起點,因此研究病毒的感染機制是了解和控制病毒的重要基礎(chǔ),已有的研究表明HPV衣殼蛋白中的堿性氨基酸區(qū)域和細(xì)胞表面硫酸乙酰肝素蛋白聚糖(HSPG)受體之間存在特異性相互作用。我們設(shè)計合成了一系列潛在的結(jié)合細(xì)胞受體的靶點多肽,利用ITC、NMR、SLS、FL及CD等實驗檢測了它們與細(xì)胞表面受體模擬物肝素之間的相互作用,并對相關(guān)作用機制進(jìn)行了系統(tǒng)研究。主要內(nèi)容包括： (一)、設(shè)計合成了HPV-16衣殼蛋白的五個多肽模擬物,檢測了它們與肝素之間的識別及相互作用。實驗結(jié)果表明：多肽結(jié)合肝素不僅受多肽本身氨基酸殘基種類和數(shù)目影響,而且存在明顯的序列特異性依賴現(xiàn)象,除最基本的靜電作用在內(nèi),疏水作用和氫鍵作用等也都參與了這些多肽與肝素的結(jié)合。另外,除了L1蛋白,L2在HPV的感染過程中也參與了結(jié)合細(xì)胞表面受體,同時證實硫酸乙酰肝素蛋白聚糖確實是細(xì)胞結(jié)合HPV的主要受體。 (二)、設(shè)計合成了HPV-16和HPV-18L1的C端相對應(yīng)位置的14肽,比較這兩個多肽結(jié)合肝素的差別。結(jié)果表明,盡管H16Ctb和H18Ctb兩個多肽所含的氨基酸種類與個數(shù)比較接近,但是因為氨基酸殘基排列順序的不同,導(dǎo)致在與肝素結(jié)合時,二者在結(jié)合能力與分子機制方面都有很大的差別。 (三)、分別設(shè)計合成了具有不同感染性質(zhì)的八種型別HPV的L1蛋白C端多肽,同樣利用ITC和SLS等方法研究了這些多肽與肝素的結(jié)合過程,確認(rèn)了它們是病毒結(jié)合細(xì)胞受體的靶點,比較了它們在結(jié)合宿主細(xì)胞受體方面的差別。 綜上所述,我們利用物理化學(xué)實驗方法研究了HPV衣殼蛋白特異性多肽與肝素之間的相互作用,定量證實了衣殼蛋白的富含堿性氨基酸區(qū)域與細(xì)胞表面硫酸乙酰肝素蛋白聚糖在病毒感染細(xì)胞中的重要作用,研究了它們之間的作用機制,在此基礎(chǔ)上為不同型別的不同感染性質(zhì)提出一種可能的解釋。
[Abstract]:Human papillomavirus (HPV) is a kind of epitheliophilic double-stranded DNA virus. Up to now, more than 100 different types of HPV have been isolated from human skin and mucosal tissues. The life cycle of a virus begins with infected cells, so studying the mechanism of virus infection is an important basis for understanding and controlling viruses. Studies have shown that there is a specific interaction between the basic amino acid region of HPV capsid protein and the heparin sulfate proteoglycan (HSPG) receptor on the cell surface. We have designed and synthesized a series of potential peptides targeting cell receptors. The interaction between these peptides and heparin, a cell surface receptor mimic, was investigated by ITCNMR-SLSS-FL and CD experiments, and the mechanism of the interaction was systematically studied. The main contents are as follows: (1) five peptide simulants of HPV-16 capsid protein were designed and synthesized, and their recognition and interaction with heparin were detected. The results showed that the peptide binding heparin was not only affected by the amino acid residues and the number of amino acids, but also had a sequence specific dependence, except for the most basic electrostatic action. Hydrophobic and hydrogen bonding are also involved in the binding of these peptides to heparin. In addition, L1 protein L2 is involved in the binding cell surface receptor in the process of HPV infection, and it is confirmed that heparin sulfate proteoglycan is the main receptor of cell binding HPV. (2) 14 peptides corresponding to C-terminal of HPV-16 and HPV-18L1 were designed and synthesized, and the difference of heparin binding between HPV-16 and HPV-18L1 was compared. The results showed that although the amino acids of H16Ctb and H18Ctb were similar to each other, the amino acid residues of H16Ctb and H18Ctb were different in order, which led to the binding with heparin. There are great differences in binding ability and molecular mechanism between them. (3) eight C-terminal peptides of L1 protein with different infection properties were designed and synthesized respectively. The binding process of these peptides to heparin was also studied by using ITC and SLS methods, which confirmed that they were the target of viral binding cell receptor. Their differences in binding to host cell receptors were compared. In conclusion, we studied the interaction between HPV capsid protein specific polypeptide and heparin by physical and chemical methods. The important role of heparin sulfate proteoglycan on the surface of capsid protein, which is rich in basic amino acids, was quantitatively confirmed, and the mechanism between them was studied. On this basis, a possible explanation for different types of infection properties is proposed.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2011
【分類號】：R373

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