基于新材料和新试剂的糖蛋白、糖肽富集和质谱鉴定新方法
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- 英文论文题名：New Materials and Reagents for Efficient Enrichment of Glycosylated Proteins and Peptides and Identification by Mass Spectrometry
- 论文作者：秦伟捷 ; 钱小红
- 英文论文作者：Weijie Qin ; Xiaohong Qian ; National Center for Protein Sciences Beijing ; State Key Lab oratory of Proteomics
- 年：2016
- 作者机构：军事医学科学院,国家蛋白质科学中心(北京);
- 论文关键词：蛋白质组学 ; 生物质谱 ; 糖肽 ; 富集 ; 定量
- 英文论文关键词：Proteomics ; Bio-mass spectrometry ; Glycopeptides ; Enrichment ; Quantification
- 会议召开时间：2016-07-01
- 会议录名称：中国化学会第30届学术年会摘要集-第四十三分会：质谱分析
- 语种：中文
- 分类号：O629.7;O657.63
- 学会代码：ZGHY
- 会议名称：中国化学会第30届学术年会-第四十三分会 ; 质谱分析
- 会议地点：中国辽宁大连
- 主办单位：中国化学会
- 学会名称：中国化学会
- 页数：1
- 文件大小：146k
- 原文格式：D
- 会议级别：全国
摘要
我们近年来发展了一系列糖蛋白、糖肽富集和标记新材料、新试剂。(1)利用高度可控的连续原子转移自由基聚合法,制备了分子结构规整的多组分嵌段亲水聚合物包裹的新型硅胶填料。巧妙整合了不同固定相特有的糖链亲和性,使该填料具有更为广谱的糖肽富集能力。(2)针对传统固态富集材料存在固液界面传质阻力和空间位阻,创新性的提出了基于可溶性环境响应材料的类均相反应富集新策略。通过调节溶液体系的温度或pH值,可对该类材料在水溶液中的溶解性实现精确控制,从而使该类材料完全溶解形成均相体系进行糖肽富集;之后使材料沉淀析出,完成样品回收并进行质谱分析。(3)发展了基于化学酶促标记的O-GlcNAc糖肽富集新方法,在O-GlcNAc糖肽上引入高甘露糖结构,从而可以使用具有更强亲和性的刀豆凝集素(ConA)对标记的O-GlcNAc糖肽进行高效富集。上述新材料、新方法显著提高了糖蛋白、糖肽的富集效率和质谱鉴定规模,为系统性的糖蛋白质组研究提供了有力的工具。
We developed a series of new materials and reagents for efficient enrichment of glycoprotein/glycopeptides.(1) We prepared sequence controlled triblock copolymer grafted silica-microparticles by sequential-ATRP of three different hydrophilic monomers.The triblock copolymer blocks carry densely packed pendent zwitterionic-ions and sugar moieties.Therefore,increased retention of glycopeptides can be achieved by the combination of different HILIC interactions.(2) We established a stimuli-responsive polymer system for facile and efficient enrichment glycopeptides.The soluble polymer supports provide a homogeneous reaction system with fast mass transfer and facilitate interactions between the supports and the target glycopeptides.More importantly,the stimuli-responsive polymers exhibit reversible self-assembly and phase separation under environmental condition variations,which leads to facial sample recovery with a high yield of the target glycopeptides.(3) We developed a new chemoenzymatic labeling based O-GlcNAc peptides enrichment method in which high mannose glycans are attached to O-GlcNAc peptides to facilitate enrichment by lectin Con A.
We developed a series of new materials and reagents for efficient enrichment of glycoprotein/glycopeptides.(1) We prepared sequence controlled triblock copolymer grafted silica-microparticles by sequential-ATRP of three different hydrophilic monomers.The triblock copolymer blocks carry densely packed pendent zwitterionic-ions and sugar moieties.Therefore,increased retention of glycopeptides can be achieved by the combination of different HILIC interactions.(2) We established a stimuli-responsive polymer system for facile and efficient enrichment glycopeptides.The soluble polymer supports provide a homogeneous reaction system with fast mass transfer and facilitate interactions between the supports and the target glycopeptides.More importantly,the stimuli-responsive polymers exhibit reversible self-assembly and phase separation under environmental condition variations,which leads to facial sample recovery with a high yield of the target glycopeptides.(3) We developed a new chemoenzymatic labeling based O-GlcNAc peptides enrichment method in which high mannose glycans are attached to O-GlcNAc peptides to facilitate enrichment by lectin Con A.
引文
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