花生致敏糖蛋白Ara h1糖链决定簇的质谱分析

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

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%.

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）,MSnand 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.