Identification and functional characterization of alpha-enolase from Taenia pisiformis metacestode

文献类型： 外文期刊

第一作者： Zhang, Shaohua

作者： Zhang, Shaohua;Guo, Aijiang;Zhu, Xueliang;You, Yanan;Hou, Junling;Luo, Xuenong;Cai, Xuepeng;Wang, Qiuxia

作者机构：

关键词： Taenia pisiformis metacestode;Enolase;Calcareous corpuscles;Plasminogen binding

期刊名称：ACTA TROPICA （ 影响因子：3.112； 五年影响因子：3.002 ）

ISSN： 0001-706X

年卷期： 2015 年 144 卷

页码：

收录情况： SCI

摘要： Enolase belongs to glycolytic enzymes with moonlighting functions. The role of enolase in Taenia species is still poorly understood. In this study, the full length of cDNA encoding for Taenia pisiformis alpha-enolase (Tpeno) was cloned from larval parasites and soluble recombinant Tpeno protein (rTpeno) was produced. Western blot indicated that both rTpeno and the native protein in excretion-secretion antigens from the larvae were recognized by anti-rTpeno monoclonal antibodies (MAbs). The primary structure of Tpeno showed the presence of a highly conserved catalytic site for substrate binding and an enolase signature motif. rTpeno enzymatic activities of catalyzing the reversible dehydration of 2-phosphoglycerate (2-PGA) to phosphoenolpyruvate (PEP) and vice versa were shown to be 30.71 +/- 2.15 U/mg (2-PGA to PEP) and 11.29 +/- 2.38 U/mg (PEP to 2-PGA), respectively. Far-Western blotting showed that rTpeno could bind to plasminogen, however its binding ability was inhibited by E-aminocaproic acid (EACA) in a competitive ELISA test. Plasminogen activation assay showed that plasminogen bound to rTpeno could be converted into active plasmin using host-derived activators. Immunohistochemistry and immunofluorescence indicated that Tpeno was distributed in the bladder wall of the metacestode and the periphery of calcareous corpuscles. In addition, a vaccine trial showed that the enzyme could produce a 36.4% protection rate in vaccinated rabbits against experimental challenges from T. pisiformis eggs. These results suggest that Tpeno with multiple functions may play significant roles in the migration, growth, development and adaptation of T. pisiformis for survival in the host environment. (C) 2015 Elsevier B.V. All rights reserved.

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