高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (9): 1622-1628.doi: 10.7503/cjcu20160147

• 分析化学 • 上一篇    下一篇

花生致敏糖蛋白Ara h1糖链决定簇的质谱分析

彭艺芳, 王承健, 王晶晶, 李玲梅, 晋万军, 强珊, 师红丹, 张英, 黄琳娟, 王仲孚()   

  1. 西部资源生物与现代生物技术教育部重点实验室, 陕西省生物技术重点实验室,西北大学生命科学学院, 西安 710069
  • 收稿日期:2016-03-14 出版日期:2016-09-10 发布日期:2016-08-26
  • 作者简介:联系人简介: 王仲孚, 男, 博士, 教授, 博士生导师, 主要从事糖生物学与糖工程研究. E-mail:wangzhf@nwu.edu.cn
  • 基金资助:
    国家自然科学基金(批准号: 21375103, 31370804)资助

Analysis of Antigenic Determinant Glycans of Peanut Allergy Glycoprotein Ara h1 by Mass Spectrometry

PENG Yifang, WANG Chengjian, WANG Jingjing, LI Lingmei, JIN Wanjun, QIANG Shan, SHI Hongdan, ZHANG Ying, HUANG Linjuan, WANG Zhongfu*()   

  1. Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education,Provincial Key Laboratory of Biotechnology, College of Life Sciences, Northwest University, Xi'an 710069, China
  • Received:2016-03-14 Online:2016-09-10 Published:2016-08-26
  • Contact: WANG Zhongfu E-mail:wangzhf@nwu.edu.cn
  • Supported by:
    † Supported by the National Natural Science Foundation of China(Nos.21375103, 31370804)

摘要:

以花生种子总蛋白及其主要致敏糖蛋白Ara h1为研究对象, 采用“一釜法”对蛋白上的糖链进行释放并同时进行衍生化标记, 通过C18固相萃取柱纯化, 以电喷雾质谱(ESI-MS)、 多级串联质谱(MSn)和亲水性液相色谱-质谱联用(HILIC-MS)进行结构解析和定量分析. 结果表明, 蛋白Ara h1共有10条N-糖链, 其中7条为高甘露糖型, 2条为木糖修饰, 另外1条为与过敏原相关的核心α1,3-Fuc修饰N-糖链, 其含量约占总糖链的12.45%.

关键词: 花生过敏, 致敏糖蛋白Ara h1, 糖链结构, 电喷雾质谱, 亲水性液相色谱-质谱联用

Abstract:

Peanut seed total protein and glycoprotein Ara h1, which can cause allergic reactions, were chosen as the target materials in this study. Firstly, the glycans were released and labeled with 1-phenyl-3-methyl-5-pyrazolone(PMP) by recently developed “one-pot” method. Subsequently, the glycans were fractionated and enriched through C18 solid phase extraction column. Finally, the obtained glycans were analysed by electrospray ionization mass spectrometry(ESI-MS), MSn and Hydrophilic interaction liquid chsomatography-mass spectrometry(HILIC-MS). The results indicated that Ara h1 has ten N-glycans, including seven high-mannose type, two ones with xylose modification, and one allergenic core α1,3-fucosylated N-glycan, the content of which is 12.45% of total glycans.

Key words: Peanut allergy, Allergenic glycoprotein Ara h1, Glycan structure, Electrospray ionization mass spectrometry(ESI-MS), Hydrophilic interaction liquid chsomatography-mass spectrometry(HILIC-MS)

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