植物凝集素串联法富集纯化血清多个亚糖蛋白质组  

Purification and Enrichment of Serum Multiple Sub-Glycoproteome by Lectins in Tandem

作　　者：齐义军[1] 王攀[1] 牛保华[1,2] 侯艳芳[1] 晁玮霞[1] WARD Douglas JOHSON Philip J 马远方[1] 

机构地区：[1]河南大学医学院细胞与分子免疫学重点实验室,开封475000 [2]河南大学环境医学研究所,开封475000 [3]CRUK-Institute for Cancer Studies,Birmingham University

出　　处：《中国科学：生命科学》2011年第5期377-383,共7页Scientia Sinica(Vitae)

基　　金：国家自然科学基金(批准号:30700366和81072039)资助项目

摘　　要：蛋白质糖基化修饰是哺乳动物中最为常见的一种翻译后修饰,蛋白质的寡糖侧链具有重要的生物学意义,如蛋白质分子间及细胞间相互作用、识别、肿瘤侵袭与转移等.本实验应用寡甘露糖型亲合层析柱、唾液酸型层析柱和O-连接糖蛋白亲合层析柱从血清中序列性提取寡甘露糖型、唾液酸型的N-连接糖蛋白及O-连接的糖蛋白,一维和二维电泳图谱显示血清各亚糖蛋白组图谱差异,并对富集的唾液酸型糖蛋白质组的二维电泳图谱中12个显著的差异蛋白质点进行质谱检测,鉴定出的14个蛋白均为N-连接糖蛋白.植物凝集素序列性提取血清各亚糖蛋白质组,显著减小了血清蛋白质组的复杂性和血清蛋白浓度波动范围,提高了质谱鉴定血清中对疾病诊断和疗效监测具有重要意义的生物标志物的灵敏性.Glycosylation is the most common protein modification post translation and sugar chains play important roles in a variety of physiological functions, e.g., recognition of proteins and cells, tumor invasion and metastasis, etc. The current study used multiple lectin chromatography in tandem to sequentially extract serum sub-glycoproteome of N-linked high-mannose and sialic acid type, and O-linked type, respectively. One-dimensional and two-dimensional SDS-PAGE showed distinct maps of each serum sub-glycoproteome. For N-linked sialic acid glycoproteome, we identified a total of 14 glycoproteins from 12 differential protein spots by ESI-MS/MS. In conclusion, sequential extraction of multiple serum glycoproteome by lectins in tandem can effectively decrease complexity and dynamic range of serum proteome, which leads to improvement of biomarker discovery for disease diagnosis and monitoring with clinical relevance.

关 键 词：糖蛋白质组 血清蛋白质组学 植物凝集素 高效液相色谱-电喷雾串联质谱 

分 类 号：Q51[生物学—生物化学]