文献类型：期刊文献

中文题名：半胱氨酸基麦芽糖修饰亲水硅胶的制备及其对糖肽的富集

英文题名：Preparation of cysteine-click maltose modified silica as a hydrophilic interaction liquid chromatography material for the enrichment of glycopeptides

作者：孙旭东[1];张凌怡[1];张维冰[1]

机构：[1]上海市功能性材料化学重点实验室,华东理工大学化学与分子工程学院,上海200237

年份：2017

卷号：35

期号：7

起止页码：696

中文期刊名：色谱

外文期刊名：Chinese Journal of Chromatography

收录：CSTPCD;;Scopus;WOS:【ESCI(收录号:WOS:000411256800004)】；北大核心:【北大核心2014】；CSCD:【CSCD2017_2018】；PubMed;

基金：National Natural Science Foundation of China (No. 21235005), National Key Scientific Instrument and Equipment Development Project (No. 2012YQ120044).

语种：中文

中文关键词：半胱氨酸;麦芽糖;硅胶;富集;糖肽;鼠肝

外文关键词：cysteine;maltose;silica;enrichment;glycopeptide;mouse liver

摘要：糖蛋白与糖肽在复杂生物体系中丰度一般较低,为了在糖蛋白质组学研究中深入和全面分析鉴定糖基化位点与糖链,通常需要进行富集前处理操作。该文设计并合成了一种半胱氨酸基麦芽糖修饰亲水硅胶分离介质(Cys-Mal@SiO_(2)),将其装填入固相萃取柱中制备新型亲水固相萃取柱。在以人免疫球蛋白G酶解液为样品进行富集鉴定时,Cys-Mal@SiO_(2)鉴定糖肽的质谱信号强度和信噪比比半胱氨酸修饰硅胶(Cys@SiO_(2))、麦芽糖修饰硅胶(Mal@SiO_(2))和商品化ZIC-HILIC更高。在复杂的鼠肝蛋白质提取物的糖蛋白质组学分析中,Cys-Mal@SiO_(2)共鉴定出1551条糖肽,属于466个糖蛋白的906个N-糖基化位点,比Cys@SiO_(2)和Mal@SiO_(2)鉴定糖肽数、蛋白质数和N-糖基化位点数分别多211、67、127个和289、76、193个。将Cys-Mal@SiO_(2)成功应用于低丰度糖肽的选择性富集与鉴定,在糖蛋白质组学研究中体现出良好的应用潜力。
Because of the low abundance of glycoprotein and glycopeptide in complex biological samples,it is urgent to develop an efficient method for glycopeptide enrichment in compre-hensive and indepth glycoproteomes research.Herein,a novel hydrophilic silica was developed through surface modification with cysteine-click maltose(Cys-Mal@SiO_(2)).The developed hydrophilic silica was packed into a solid phase extraction(SPE)column,and applied to the highly selective enrichment and identification of Hinked glycopeptides.The Cys-Mal@SiO_(2) demonstrated better identification capability over Cys@SiO_(2),Mal@SiO_(2) and commercial hydro-philic interaction liquid chromatography(HILIC)in glycopeptide enrichment due to the syner-gistic effect of the two kinds of hydrophilic molecules.In the selective enrichment of tryptic digest from human immunoglobulin G,glycopeptides with higher signal-to-noises were detected by Cys-Mal@SiO2.In addition,1551 unique glycopeptides with 906 N-glycosylation sites from 466 different N-linked glycoproteins were identified from the proteins extracted from mouse liver after the enrichment with Cys-Mal@SiO_(2).In contrast,the numbers of identified glycopeptides,glycoproteins and N-glycosylation sites identified by Cys@SiO_(2) were 211,67,127 respectively less than by Cys-Mal@SiO_(2),and the corresponding numbers were 289,76,193 by Mal@SiO_(2).These results showed that the developed Cys-Mal@SiO_(2) is a promising affinity material for N-glycoproteomics research of real complex biological samples.