凝集素法与酰肼法富集糖肽的方法学比较  

Comparison of Hydrazide Chemistry and Lectin Affinity Based Enrichment Methods for N-glycoproteomics

作　　者：薛鹏[1,2] 周岳[1,2,3] 丁翔[1] 王峻[1,2] 谢振声[1] 杨福全[1] 

机构地区：[1]中国科学院生物物理研究所,蛋白质组学技术实验室,北京100101 [2]中国科学院大学,北京100049 [3]赛默飞世尔科技有限公司,上海200120

出　　处：《生物化学与生物物理进展》2015年第4期356-364,共9页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展计划(973)(2012CB966803,2010CB833703);中国科学院仪器设备功能开发技术创新项目(yg201001)资助~~

摘　　要：蛋白质糖基化(glycosylation)是最常见和最重要的翻译后修饰之一.大规模N-连接糖基化位点鉴定是糖蛋白质组学研究的重要组成部分,而N-连接糖肽富集是高通量N-连接糖基化位点鉴定的关键步骤.凝集素富集法和酰肼化学法是目前被广泛应用的N-连接糖肽富集技术,有报道认为两种方法具有很强的互补性,联合使用能提高糖基化位点的鉴定数目.本文以Hep G2细胞系为模型,系统比较了这两种方法的富集效率和糖蛋白鉴定数目.结果表明,虽然酰肼法的糖肽富集效率为76.6%,远高于凝集素法的54.6%,但是凝集素法却能鉴定到825个糖蛋白和1 959个N-连接糖基化位点,显著多于酰肼法富集到的522个糖蛋白和1 014个糖基化位点.并且,两种方法并未显示出显著的互补性,仅28个糖蛋白和80个糖基化位点未在凝集素法中鉴定到.Glycosylation is one of the most common and important post-translational modifications of proteins. Identification of large-scale N-linked glycoprotein is a very important aspect in glycoproteomics research. The N-glycopeptide enrichment is a key step in high-throughput identification of N-glycosylation site. Lectin enrichment and hydrazide chemistry are the two widely used N-glycopeptides enrichment methods. Each method can only enrich certain types of glycopeptides. It has been reported that the two methods are highly complementary, but few studies compared overlaps of glycosties from the two methods. In this paper, using HepG2 cells, we systematically compared the performance of hydrazide chemistry and lectins enrichment methods. The results showed that although the hydrazide method with glycopeptides enrichment efficiency of 76.7%, far higher than the 54.6% lectin enrichment method, 825 glycoprotein and 1 879 N-glycosylation sites identified with the lectin method was significantly more than 522 glycoprotein and 1 014 glycosylation sites enriched by the hydrazide method. Moreover, the two methods did not show significant complementary, together, only 853 glycoproteins and 1 959 N-glycosylation sites were identified. The overlapping results of identified N-glycosylation sites and N-glycoproteins from the two methods show that leetins enrichment method was better than hydrazide chemistry method.

关 键 词：N-连接糖基化蛋白质组学 凝集素富集法 酰肼化学富集法 

分 类 号：O658[理学—分析化学] Q51[理学—化学]