Proteome analysis of Clonorchis sinensis

초록

Clonorchiasis is a hepatobiliary disease caused by liver fluke, Clonorchis sinensis and is one of the important parasitic diseases in Korea. In spite of advances in investigating the immunologic molecules of C. sinensis, there are still lacking for reliable diagnostic and prognostic markers in clonorchiasis. In this study, we have established 2-dimensional proteome map and identified diagnostic candidates useful for serodiagnosis of clonorchiasis employing immunoproteomics. The total protein map of ESP (excretory-secretory product) and total lysates was constructed by using 2-DE in the range of pI 3 -11 followed by immuno-blot against clonorchiasis patients’ sera. We identified 62 in ESP and 102 protein spots in total lysates, respectively, by using mass analysis and expressed sequence tag (EST) database of C. sinensis. The most prominent proteins were identified as proteases, myoglobin and glutathione transferase related to peptidase activity, oxygen transport and detoxification, respectively. Analysis of glycoproteins by lectin blotting employing Canavalia ensiformis (Con-A) and Helix pomatia (HPA) found a variety of glycoproteins of a broad molecular mass from 10 to 120 kDa and pI range from 3 to 11. The comparative immunoglycomic analysis revealed that several antigenic proteins against patient sera were overlapped on glycoprotein spots on Con-A lectin blot, but not on that of HPA. This result suggested that most of antigenic proteins in C. sinensis were high mannose type N-linked glycoproteins. Identified proteins are valuable to be further investigations, which may start for a new research on immunological action and host interaction in clonorchiasis.