Sequence analysis and characterization of pyruvate kinase from Clonorchis sinensis, a 53.1-kDa homopentamer, implicated immune protective efficacy against clonorchiasis

Parasites & Vectors

卷:10

Pengli Ren¹  Zhizhi Xie²  Xinbing Yu³  Yan Huang³  Mengchen Shi³  Zhiyue Lv³  Jin Xu³  Xuerong Li³  Hengchang Sun³  Huimin Dong³  Lu Zhao³  Tingjin Chen³  Hongye Jiang³  Zhongdao Wu³ 

[1] Department of Clinical Laboratory, Second Affiliated Hospital of Guangzhou Medical University;

[2] Department of Clinical Laboratory, Third Affiliated Hospital, Sun Yat-sen University;

[3] Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University;

关键词: Clonorchis sinensis;    Pyruvate kinase;    Pentamer;    Expression profile;    Excretory/secretory products;    Immune response;   

DOI  :  10.1186/s13071-017-2494-9

来源: DOAJ

【 摘 要 】

Abstract Background Clonorchis sinensis, the causative agent of clonorchiasis, is classified as one of the most neglected tropical diseases and affects more than 15 million people globally. This hepatobiliary disease is highly associated with cholangiocarcinoma. As key molecules in the infectivity and subsistence of trematodes, glycolytic enzymes have been targets for drug and vaccine development. Clonorchis sinensis pyruvate kinase (CsPK), a crucial glycolytic enzyme, was characterized in this research. Results Differences were observed in the sequences and spatial structures of CsPK and PKs from humans, rats, mice and rabbits. CsPK possessed a characteristic active site signature (IKLIAKIENHEGV) and some unique sites but lacked the N-terminal domain. The predicted subunit molecular mass (Mr) of CsPK was 53.1 kDa. Recombinant CsPK (rCsPK) was a homopentamer with a Mr. of approximately 290 kDa by both native PAGE and gel filtration chromatography. Significant differences in the protein and mRNA levels of CsPK were observed among four life stages of C. sinensis (egg, adult worm, excysted metacercaria and metacercaria), suggesting that these developmental stages may be associated with diverse energy demands. CsPK was widely distributed in adult worms. Moreover, an intense Th1-biased immune response was persistently elicited in rats immunized with rCsPK. Also, rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. Conclusions The sequences and spatial structures, molecular mass, and expression profile of CsPK have been characterized. rCsPK was indicated to be a homopentamer. Rat anti-rCsPK sera suppressed C. sinensis adult subsistence both in vivo and in vitro. CsPK is worthy of further study as a promising target for drug and vaccine development.

【 授权许可】

Unknown