本文選題：泛素、類泛素蛋白 + 蛋白質(zhì)化學(xué)合成��； 參考：《合肥工業(yè)大學(xué)》2017年碩士論文

【摘要】：蛋白質(zhì)的泛素化及類泛素化修飾在真核生物細(xì)胞內(nèi)廣泛存在并參與了大部分重要的細(xì)胞生命過程,包括信號(hào)傳導(dǎo)、DNA損傷修復(fù)、蛋白降解等。不同底物的泛素及類泛素化修飾類型多種多樣且具有不同的功能,保證了修飾功能的多樣性和特異性。目前有相當(dāng)一部分具有重要功能的泛素及類泛素蛋白由于缺乏特異性的酶促合成系統(tǒng)而不易獲取。同時(shí),獲取純凈均一的泛素探針用于研究去泛素化酶的水解機(jī)制更為困難。蛋白質(zhì)化學(xué)合成技術(shù)憑借其能夠在原子尺度精準(zhǔn)構(gòu)筑蛋白質(zhì)的優(yōu)勢(shì),在傳統(tǒng)生物酶法獲取泛素、類泛素蛋白存在制約的情況下,提供了一種行之有效的策略。本文研究工作以蛋白質(zhì)化學(xué)合成技術(shù)為工具,圍繞泛素及類泛素蛋白的化學(xué)合成開展研究。首先通過使用本課題組發(fā)展的一鍋法兩片段連接-脫硫策略,我們高效地合成了類泛素化蛋白NEDD8以及泛素探針Ub-AMC,與前人方法相比合成步驟更少,效率更高。此后,作者又運(yùn)用基于非天然氨基酸定點(diǎn)嵌入的蛋白質(zhì)半合成方法對(duì)多聚泛素鏈的合成進(jìn)行了探索。我們首先通過嵌入Alloc-Lys并使用Boc保護(hù)其他賴氨酸側(cè)鏈氨基的方法成功制備了可用于連接的泛素片段;此后運(yùn)用輔基介導(dǎo)的自然化學(xué)連接反應(yīng)來實(shí)現(xiàn)泛素片段間異肽鍵的高效制備。本研究工作中成功制備的賴氨酸48位(K48)形成異肽鍵的二泛素驗(yàn)證了上述策略的可行性。本工作為進(jìn)一步化學(xué)合成其他位點(diǎn)的多聚泛素、類泛素鏈提供了一種普適性方法,進(jìn)而為闡明蛋白質(zhì)泛素、類泛素化修飾的特異性識(shí)別、降解機(jī)制提供基礎(chǔ)。
[Abstract]:Protein ubiquitization and ubiquitin modification exist widely in eukaryotic cells and participate in most important cellular life processes, including signal transduction DNA damage repair, protein degradation and so on. The types of ubiquitin and ubiquitin modified by different substrates are various and have different functions, which ensures the diversity and specificity of the modification functions. At present, a considerable number of ubiquitin and ubiquitin like proteins with important functions are difficult to obtain due to the lack of specific enzymatic synthesis system. At the same time, it is more difficult to obtain pure and uniform ubiquitin probe to study the hydrolysis mechanism of diubiquitin. Because protein chemical synthesis technology has the advantage of accurately constructing proteins on atomic scale, it provides an effective strategy for obtaining ubiquitin by traditional biological enzyme method and the restriction of ubiquitin protein. In this paper, the chemical synthesis of ubiquitin and ubiquitin protein was studied by using protein chemical synthesis technology as a tool. Firstly, we synthesized ubiquitin NEDD8 and Ub-AMC efficiently by using the one-pot two-fragment connection-desulphurization strategy developed by our team. Compared with previous methods, we synthesized Ub-AMCs with less steps and higher efficiency. Since then, the author has explored the synthesis of polyubiquitin chain by protein semi-synthesis method based on non-natural amino acid fixed point intercalation. Ubiquitin fragments were successfully prepared by embedding Alloc-Lys and protecting other lysine side chain amino groups by Boc, and then the heteropeptide bonds between Ubiquitin fragments were efficiently prepared by natural chemical bonding mediated by auxiliary groups. The diubiquitin of isopeptide bond formed by Lysine 48 K48 was proved to be feasible. This work provides a universal method for the further chemical synthesis of polyubiquitin and ubiquitin chain at other sites and provides a basis for elucidating the specific recognition and degradation mechanism of protein ubiquitin and ubiquitin modification.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O629.73

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 LIANG Jun;FANG GeMin;HUANG XiuLiang;MEI ZiQing;LI Juan;TIAN ChangLin;LIU Lei;;Chemical synthesis of Ub-AMC via ligation of peptide hydrazides[J];Science China(Chemistry);2013年09期

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本文編號(hào)：1778137