N-连接完整糖肽的质谱分析策略和研究方法
发布时间：2018-07-17 00:06
【摘要】：蛋白质糖基化作为最普遍、最重要的蛋白质修饰,一直是组学研究的焦点之一.近十几年来,N-连接糖蛋白质组学研究普遍采用的方法是将糖链与所修饰的多肽分开进行分析.该策略虽降低了分析难度,却也丢失了糖链与蛋白质糖基化位点间重要的对应关系信息.近年来,完整糖肽的质谱分析策略和方法逐步建立起来.总体而言,要实现对完整糖肽的直接质谱分析,首先需要从复杂样品中富集完整糖肽以消除非糖基化多肽对完整糖肽分析的影响,然后在质谱分析中还需要根据糖肽特性调整相应质谱分析参数,最后在后续数据分析中还需要开发相应的分析软件以完成完整糖肽中多肽序列和糖链组成或结构的鉴定.本文即从以上三个主要方面系统阐述目前N-完整糖肽分析中常用的质谱和数据分析策略和方法,并进一步在糖肽谱图识别、母离子单同位素分子质量校正、数据库选择以及假阳性率评估和控制等方面都进行了逐一探讨.完整糖肽的直接质谱分析有助于获取糖链和糖基化位点间的对应关系信息,可为生物标志物发现和疾病致病机理等研究提供更有力的糖蛋白质组学研究工具.
[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.
【作者单位】： 西北大学生命科学学院;
【基金】：中组部“青年千人”配套经费(361010001)资助~~
【分类号】：O629.72;O657.63
[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.
【作者单位】： 西北大学生命科学学院;
【基金】：中组部“青年千人”配套经费(361010001)资助~~
【分类号】：O629.72;O657.63
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