Identification and molecular characterization of a novel signaling molecule 14-3-3 epsilon in Clonorchis sinensis excretory/secretory products

• Published in: Parasitology research (1987) Vol. 110; no. 4; pp. 1411 - 1420
• Main Authors: Wang, Xiaoyun, Chen, Wenjun, Li, Xuerong, Zhou, Chenhui, Deng, Chuanhuan, Lv, Xiaoli, Fan, Yongxiu, Men, Jingtao, Liang, Chi, Yu, Xinbing
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01.04.2012; Springer
• Subjects: 14-3-3 Proteins - genetics; 14-3-3 Proteins - immunology; 14-3-3 Proteins - isolation & purification; 14-3-3 Proteins - metabolism; Amino Acid Sequence; Animals; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Cloning, Molecular; Clonorchiasis - diagnosis; Clonorchiasis - parasitology; Clonorchis sinensis - genetics; Clonorchis sinensis - immunology; Clonorchis sinensis - pathogenicity; Computational Biology; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling; General aspects; General aspects and techniques. Study of several systematic groups. Models; Helminth Proteins - genetics; Helminth Proteins - immunology; Helminth Proteins - isolation & purification; Helminth Proteins - metabolism; Humans; Immunoblotting; Immunoglobulin G - blood; Immunology; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Invertebrates; Medical Microbiology; Metacercariae - genetics; Metacercariae - immunology; Microbiology; Molecular dynamics; Molecular Sequence Data; Original Paper; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Rats; Rats, Sprague-Dawley; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sensitivity and Specificity; Sequence Analysis, DNA; Signal Transduction; Th1-Th2 Balance; Vaccination; Virulence (Microbiology); China; Adult Worm; Clonorchiasis; Schistosomiasis; Cysticercosis; Fascioliasis; Characterization; Molecules; Helmintha; Plathelmintha; Parasite; Clonorchis sinensis; Genetic identification; Invertebrata; Trematoda
• ISSN: 0932-0113; 1432-1955
• DOI: 10.1007/s00436-011-2642-7

Increasing evidence shows that 14-3-3 proteins are involved in many biology events in addition to signal transduction. Extensive investigations on structural and biochemical features of these signaling molecules have implied their importance in the biological process. In the present study, we have identified and characterized the 14-3-3 epsilon ( Cs 14-3-3) in Clonorchis sinensis that causes human clonorchiasis. Recombinant protein was expressed in Escherichia coli ( E. coli ) and identified by MALDI-TOF/TOF. Immunoblot results revealed that Cs 14-3-3 was a component of excretory/secretory products. Ligand blot assay indicated that 14-3-3 epsilon could bind C. sinensis MAPKAPK 2 in a nonphosphorylation-dependent manner. This protein could be detected at four stages of the life cycle by RT-PCR experiments and immunolocalization showed that Cs 14-3-3 was extensively distributed in C. sinensis , especially at the outer surface and the sucker of adult worm and cyst wall of metacercaria. Taken together, 14-3-3 epsilon might play some roles in the development of the parasites. In addition, Cs 14-3-3 epsilon should be addressed for the diagnostic value in C. sinensis infection in consideration of high sensitivity and specificity. As an immune stimulus, C. sinensis 14-3-3 epsilon was found to provoke a Th1/Th2 balanced immune response by inducing high levels of both IgG1 and IgG2a. Recombinant Cs 14-3-3 conferred effective protection both in worm reduction rate and egg reduction rate, suggesting that the signaling molecule Cs 14-3-3 was a promising vaccine candidate against C. sinensis infection.