Abstract
Tegumental proteins form a membrane-bound outer surface and are thus involved in host–parasite interactions and parasite survival. A complementary DNA clone encoding a novel 21.6-kDa tegumental protein (CsTegu21.6, accession number JF911532) was identified in a sequence library for the adult Clonorchis sinensis liver fluke. The complete coding sequence was 564 bp and encoded a protein of 188 amino acids. A BLASTX search revealed identities from 43 to 47% with previously identified tegumental proteins in C. sinensis and other helminthic parasites. Multiple alignment of the amino acids of CsTegu21.6 with those of four other C. sinensis tegumental proteins, CsTegu21.1, CsTegu22.3, CsTegu20.8 and CsTegu31.8, revealed pair-wise sequence identities ranging from 24 to 31.8%. A calcium-binding EF-hand domain containing a basic helix–loop–helix structure at the N terminus and a dynein light chain domain at the C terminus were found in CsTegu21.6; these motifs are common in tegumental proteins. CsTegu21.6 was specifically observed on the tegument of adult worms using immunolocalization analysis.
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Acknowledgments
This work was supported by a grant from the Pathogenic Proteome Management Program of the National Institute of Health, Ministry of Health and Welfare, Republic of Korea (NIH 48004847–300).
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The authors have declared that no competing interests exist.
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Yu-Jung Kim and Won Gi Yoo contributed equally to this study.
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Supplementary Figure 1
The sequence and structure of CsTegu21.6. a The cDNA and deduced amino acid sequences of CsTegu21.6 are shown. The shaded fragment indicates the EF-hand domain, and the underlined fragment represents the dynein light chain domain. The four B-cell epitopes are displayed in bold. b The 3D-structure of CsTegu21.6 was predicted using SWISS-MODEL and was visualized using PyMOL. The epitopes are colored in red (PPT 432 kb)
Supplementary Figure 2
Expression and purification of rCsTegu21.6. rCsTegu21.6 expression in E. coli BL21(DE3) was verified by 12% SDS–PAGE followed by Coomassie blue staining and western blot analysis with an anti-polyhistidine antibody. Lane 1, molecular weight marker; lane 2, E. coli prior to induction; lane 3, E. coli induced with IPTG; lane 4, flow through; lanes 5 and 6, purified recombinant proteins (PPT 244 kb)
Supplementary Table 1
LC–MS/MS identification of rCsTegu21.6 (DOC 31 kb)
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Kim, YJ., Yoo, W.G., Lee, MR. et al. Identification and characterization of a novel 21.6-kDa tegumental protein from Clonorchis sinensis . Parasitol Res 110, 2061–2066 (2012). https://doi.org/10.1007/s00436-011-2681-0
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DOI: https://doi.org/10.1007/s00436-011-2681-0