Mass spectrometric analysis of the immunodominant glycan epitope of Echinococcus granulosus antigen Ag5.

Mass spectrometric analysis of the immunodominant glycan epitope of Echinococcus granulosus antigen Ag5.
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DOI:
10.1016/j.ijpara.2012.01.002
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发表时间:
2012
影响因子:
4
通讯作者:
Wilson IB
Wilson IB
中科院分区:
医学2区
文献类型:
--
作者:
Paschinger K;Gonzalez-Sapienza GG;Wilson IB

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►细粒棘球蚴抗原Ag 5具有免疫显性聚糖表位。对E.用质谱法分析颗粒体Ag 5 38 kDa亚基。分析证实了在绦虫蛋白质上存在磷酸胆碱。在以前的工作中,我们表明,Ag 5,一个主要的诊断抗原从细粒棘球绦虫的后绦虫,具有一个显性糖表位,去除后的结果在废除大部分的抗原免疫反应性与患者血清。现在已经通过蛋白质印迹法和质谱法对这种聚糖修饰进行了分析。对特异性单克隆抗体(TEPC 15)和人C-反应蛋白的反应性以及165质量单位的修饰的存在,如通过糖肽和释放的N-聚糖的质谱法检测到的,表明Ag 5 38 kDa亚基的免疫显性糖表位是由磷酸胆碱修饰的双触角结构。我们相信这是第一次在绦虫中证明这种修饰,并为理解这种主要E.颗粒体成分
► Echinococcus granulosus antigen, Ag5, possesses an immunodominant glycan epitope. ► The N-glycans and glycopeptides of the E. granulosus Ag5 38 kDa subunit were analysed by mass spectrometry. ► Analyses verified the presence of phosphorylcholine on a cestode protein. In previous work we showed that Ag5, a major diagnostic antigen from the metacestode of Echinococcus granulosus, possesses a dominant sugar epitope that upon removal results in abolition of most of the antigen immunoreactivity with patient sera. Analysis of this glycan modification has now been performed by western blotting and mass spectrometry. Reactivity to both a specific monoclonal antibody (TEPC15) and human C-reactive protein as well as the presence of a modification of 165 mass units, as detected by mass spectrometry of both glycopeptides and released N-glycans, indicated that the immunodominant sugar epitope of the Ag5 38 kDa subunit is a biantennary structure modified by phosphorylcholine. We believe this is the first time that such a modification has been proven in cestodes and provides the structural basis for understanding the antigenicity of this major E. granulosus component.
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