Identification and characterization of an immunogenic antigen, enolase 2, among excretory/secretory antigens (ESA) of Toxoplasma gondii

文献类型： 外文期刊

第一作者： Jiang, Wei

作者： Jiang, Wei;Xue, Jun-Xin;Liu, Ying-Chun;Li, Tao;Han, Xian-Gan;Wang, Shao-Hui;Chen, Yong-Jun;Qi, Jingjing;Yu, Sheng-Qing;Wang, Quan;Xue, Jun-Xin

作者机构：

关键词： Toxoplasma gondii;Excretory/secretory antigens;Immunoproteomic;Enolase 2;Enzyme activity

期刊名称：PROTEIN EXPRESSION AND PURIFICATION （ 影响因子：1.65； 五年影响因子：1.548 ）

ISSN：

年卷期：

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收录情况： SCI

摘要： An immunogenic protein, enolase 2, was identified among the secreted excretory/secretory antigens (ESAs) from Toxoplasma gondii strain RH using immunoproteomics based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Enolase 2 was cloned, sequenced, and heterologously expressed. BLAST analysis revealed 75-96% similarity with enolases from other parasites. Immunoblotting demonstrated good immunoreactivity of recombinant T gondii enolase (Tg-enolase 2) to T. gondii-infected animal serum. Purified Tg-enolase 2 was found to catalyze dehydration of 2-phospho-D-glycerate to phosphoenolpyruvate. In vitro studies revealed maximal activity at pH 7.5 and 37 degrees C, and activity was inhibited by K+, Ni2+, Al3+, Na+, Cu2+ and Cr3+. A monoclonal antibody against Tg-enolase 2 was prepared, 1D6, with the isotype IgG2a/k. Western blotting revealed that 1D6 reacts with Tg-enolase 2 and native enolase 2, present among T. gondii ESAs. The indirect immunofluorescence assays showed that enolase 2 could be specifically detected on the growing T gondii tachyzoites. Immunoelectron microscopy revealed the surface and intracellular locations of enolase 2 on T. gondii cells. In conclusion, our results clearly show that the enzymatic activity of T. gondii enolase 2 is ion dependent and that it could be influenced by environmental factors. We also provide evidence that enolase 2 is an important immunogenic protein of ESAs from T. gondii and that it is a surface-exposed protein with strong antigenicity and immunogenicity. Our findings indicate that enolase 2 could play important roles in metabolism, immunogenicity and pathogenicity and that it may serve as a novel drug target and candidate vaccine against T gondii infection. (C) 2016 Elsevier Inc. All rights reserved.

分类号： Q51

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