【摘要】：蛋白質(zhì)糖基化作為最普遍、最重要的蛋白質(zhì)修飾,一直是組學(xué)研究的焦點(diǎn)之一.近十幾年來(lái),N-連接糖蛋白質(zhì)組學(xué)研究普遍采用的方法是將糖鏈與所修飾的多肽分開(kāi)進(jìn)行分析.該策略雖降低了分析難度,卻也丟失了糖鏈與蛋白質(zhì)糖基化位點(diǎn)間重要的對(duì)應(yīng)關(guān)系信息.近年來(lái),完整糖肽的質(zhì)譜分析策略和方法逐步建立起來(lái).總體而言,要實(shí)現(xiàn)對(duì)完整糖肽的直接質(zhì)譜分析,首先需要從復(fù)雜樣品中富集完整糖肽以消除非糖基化多肽對(duì)完整糖肽分析的影響,然后在質(zhì)譜分析中還需要根據(jù)糖肽特性調(diào)整相應(yīng)質(zhì)譜分析參數(shù),最后在后續(xù)數(shù)據(jù)分析中還需要開(kāi)發(fā)相應(yīng)的分析軟件以完成完整糖肽中多肽序列和糖鏈組成或結(jié)構(gòu)的鑒定.本文即從以上三個(gè)主要方面系統(tǒng)闡述目前N-完整糖肽分析中常用的質(zhì)譜和數(shù)據(jù)分析策略和方法,并進(jìn)一步在糖肽譜圖識(shí)別、母離子單同位素分子質(zhì)量校正、數(shù)據(jù)庫(kù)選擇以及假陽(yáng)性率評(píng)估和控制等方面都進(jìn)行了逐一探討.完整糖肽的直接質(zhì)譜分析有助于獲取糖鏈和糖基化位點(diǎn)間的對(duì)應(yīng)關(guān)系信息,可為生物標(biāo)志物發(fā)現(xiàn)和疾病致病機(jī)理等研究提供更有力的糖蛋白質(zhì)組學(xué)研究工具.
[Abstract]:Glycosylation of proteins is one of the most common and important protein modifications. In recent years, the commonly used method in the proteomics of N-ligand is to separate the sugar chain from the modified polypeptide. Although this strategy reduces the difficulty of analysis, it also loses the important correspondence information between sugar chain and glycosylation site of protein. In recent years, the strategies and methods for the analysis of complete glycopeptide by mass spectrometry have been gradually established. In general, direct mass spectrometry of complete glycopeptides requires enrichment of complete glycopeptides from complex samples to eliminate the influence of non-glycosylated peptides on the analysis of complete glycopeptides. Then we need to adjust the parameters according to the characteristics of glycopeptide in the mass spectrum analysis. Finally, in the subsequent data analysis, we also need to develop the corresponding analysis software to complete the identification of peptide sequence and sugar chain composition or structure in the complete glycopeptide. In this paper, the strategies and methods of mass spectrometry and data analysis commonly used in the analysis of N-complete glycopeptide are systematically described from the above three main aspects, and further, the identification of glycopeptide spectrum and the molecular mass correction of mother ion single isotope are carried out. The selection of database and the evaluation and control of false positive rate were discussed one by one. Direct mass spectrometry analysis of complete glycopeptides is helpful to obtain the corresponding information between glycosylation sites and glycosylation sites, and can provide a more powerful tool for the study of glycosylproteomics for the discovery of biomarkers and the pathogenesis of diseases.
【作者單位】： 西北大學(xué)生命科學(xué)學(xué)院;
【基金】：中組部“青年千人”配套經(jīng)費(fèi)(361010001)資助~~
【分類號(hào)】：O629.72;O657.63

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