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Vibrio parahaemolyticus enolase is an adhesion-related factor that binds plasminogen and functions as a protective antigen

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Abstract

Vibrio parahaemolyticus, an emerging food and waterborne pathogen, is a leading cause of seafood poisoning worldwide. Surface proteins can directly participate in microbial virulence by facilitating pathogen dissemination via interactions with host factors. Screening and identification of protective antigens is important for developing therapies against V. parahaemolyticus infections. Here, we systematically characterized a novel immunogenic enolase of V. parahaemolyticus. The enolase gene of V. parahaemolyticus ATCC33847 was cloned, sequenced, and expressed in Escherichia coli BL21. Enzymatic assays revealed that the purified recombinant V. parahaemolyticus enolase protein catalyzes the dehydration of 2-phospho-d-glycerate to phosphoenolpyruvate. Western blot analysis showed that V. parahaemolyticus enolase was detectable in the extracellular, outer membrane (OM) and cytoplasmic protein fractions using antibodies against the recombinant enolase. Surface expression of enolase was further confirmed by immunogold staining and mass spectrometry (liquid chromatography–tandem mass spectrometry) analysis of OM protein profiles. Notably, V. parahaemolyticus enolase was identified as a human plasminogen-binding protein with the enzyme-linked immunosorbent assay. The values obtained for adherence and inhibition suggest a role of surface-exposed enolase in epithelial adherence of V. parahaemolyticus. We further showed that enolase confers efficient immunity against challenge with a lethal dose of V. parahaemolyticus in a mouse model. To our knowledge, this is the first study to demonstrate the plasminogen-binding activity of enolase that is an adhesion-related factor of V. parahaemolyticus. Our findings collectively imply that enolase plays important roles in pathogenicity, supporting its utility as a novel vaccine candidate against V. parahaemolyticus infection.

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Acknowledgments

This work was funded by the National Key Technology Support Program (2012BAK08B07), Shanghai Municipal Science and Technology Commission technical standards (13DZ0502702), and Aquatic Three New Projects in Jiangsu Province (D2013-5-4).

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Authors and Affiliations

  1. College of Veterinary Medicine, Nanjing Agricultural University, 1 Weigang, Nanjing, 210095, Jiangsu, China

    Wei Jiang, Xintong Li, Li Yi & Yongjie Liu

  2. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, People’s Republic of China

    Wei Jiang, Xiangan Han, Quan Wang & Chan Ding

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  1. Wei Jiang
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  2. Xiangan Han
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  3. Quan Wang
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  4. Xintong Li
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Correspondence to Yongjie Liu or Chan Ding.

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Jiang, W., Han, X., Wang, Q. et al. Vibrio parahaemolyticus enolase is an adhesion-related factor that binds plasminogen and functions as a protective antigen. Appl Microbiol Biotechnol 98, 4937–4948 (2014). https://doi.org/10.1007/s00253-013-5471-z

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