title: Identification and molecular characterization of a novel signaling molecule 14-3-3 epsilon in Clonorchis sinensis excretory/secretory products.
abstract: 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 (Cs14-3-3) 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 Cs14-3-3 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 Cs14-3-3 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, Cs14-3-3 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 Cs14-3-3 conferred effective protection both in worm reduction rate and egg reduction rate, suggesting that the signaling molecule Cs14-3-3 was a promising vaccine candidate against C. sinensis infection.
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Chen
authors.forename: Xuerong
authors.initials: X
authors.lastname: Li
authors.forename: Chenhui
authors.initials: C
authors.lastname: Zhou
authors.forename: Chuanhuan
authors.initials: C
authors.lastname: Deng
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Lv
authors.forename: Yongxiu
authors.initials: Y
authors.lastname: Fan
authors.forename: Jingtao
authors.initials: J
authors.lastname: Men
authors.forename: Chi
authors.initials: C
authors.lastname: Liang
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D048948:14-3-3 Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D003003:Clonorchiasis / Q000175:diagnosis / Q000469:parasitology; D003004:Clonorchis sinensis / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D019295:Computational Biology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000097:blood; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D058488:Metacercariae / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D058408:Th1-Th2 Balance; D014611:Vaccination; D000099190:MAP-Kinase-Activated Kinase 2
chemical_list: D048948:14-3-3 Proteins; D015801:Helminth Proteins; D007074:Immunoglobulin G; D047908:Intracellular Signaling Peptides and Proteins; D017346:Protein Serine-Threonine Kinases; D011994:Recombinant Proteins; D000099190:MAP-Kinase-Activated Kinase 2