| 基于固相萃取的微量糖蛋白糖基化修饰表征技术的建立 |
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| 引用本文： | 江静,韩欢欢,马成,王继峰,应万涛,钱小红.基于固相萃取的微量糖蛋白糖基化修饰表征技术的建立[J].分析化学,2012(7):1019-1024. |
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| 作者姓名： | 江静 韩欢欢 马成 王继峰 应万涛 钱小红 |
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| 作者单位： | 蛋白质组学国家重点实验室,北京蛋白质组研究中心,军事医学科学院放射与辐射医学研究所;中国人民解放军总医院,军医进修学院;蛋白质药物国家工程研究中心 |
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| 基金项目： | 国家973课题(No.2011CB910603);国家自然科学基金(No.2073505,30900258);重点实验室自主研究课题(No.SKLP-K200902)资助 |
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| 摘 要： | 结合自制亲水固相萃取富集柱和生物质谱鉴定技术,实现了糖基化蛋白质核糖核酸酶B的糖含量测定、糖基化位点确认、聚糖富集及结构表征,以及不同糖型相对丰度分析。结果表明:其糖含量8.47%,糖基化位点为34位的Asn,糖链主要为5种高甘露糖型结构(Man5-9GlcNAc2)。所建立的HILIC富集技术,有利于针对微量生物样本,如生物工程药物糖蛋白及重要功能糖蛋白,开展位点特异性糖链结构解析,为糖蛋白质的药效或功能研究提供线索。
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| 关 键 词： | 糖蛋白 亲水固相萃取 基质辅助激光解吸电离飞行时间质谱 线性离子阱傅里叶变换离子回旋共振质谱仪 |
Characterization of N-Glycosylation Status of Micro-level Glycoprotein Using Solid Phase Extraction Based Techniques |
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| JIANG Jing,HAN Huan-Huan,MA Cheng,WANG Ji-Feng,YING Wan-Tao,QIAN Xiao-Hong.Characterization of N-Glycosylation Status of Micro-level Glycoprotein Using Solid Phase Extraction Based Techniques[J].Chinese Journal of Analytical Chemistry,2012(7):1019-1024. |
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| Authors: | JIANG Jing HAN Huan-Huan MA Cheng WANG Ji-Feng YING Wan-Tao QIAN Xiao-Hong |
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| Institution: | 1,3 1(State Key Laboratory of Proteomics,Beijing Proteome Research Center,Beijing Institute of Radiation Medicine,Beijing 102206,China) 2(Chinese People’s Liberation Army General Hospital,Chinese People′s Liberation Army Postgraduate Medical School,Beijing 100039,China) 3(National Engineering Research Center for Protein Drugs,Beijing 102206,China) |
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| Abstract: | A new strategy integrating the latest hydrophilic interaction chromatographic(HILIC) method with mass spectrometry technologies was developed for the enrichment and determination of intact glycopeptides and glycan structures from a glycoprotein.The average molecular weight,the ratio of glycosylation,the glycosite and the structures of glycoforms of ribonuclease B(RNaseB) were determined by MALDI-TOF-MS and ESI-LTQ-FT-MS.RNaseB contains 8.43% glycan and has a single glycosylation site at Asn34 with the glycan structure varied from five to nine mannose residues(Man5-9GlcNAc2).The strategy paves the way for systematic and confident analysis of complex glycoproteins with important biological or pharmaceutical functions. |
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| Keywords: | Glycoprotein Hydrophilic interaction chromatographic solid phase extraction Matri-xassisted laser desorption/ionization-time of flight-mass spectrometry Linear quadrupole ion trap-Fourier transform ion cyclotron resonance mass spectrometry |
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