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Primary structure, expression and localization of two intermediate subunit lectins of Entamoeba dispar that contain multiple CXXC motifs

Published online by Cambridge University Press:  06 September 2007

H. TACHIBANA ,
X.-J. CHENG ,
S. KOBAYASHI ,
Y. OKADA ,
J. ITOH  and
T. TAKEUCHI
H. TACHIBANA*
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
X.-J. CHENG
Affiliation:
Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
S. KOBAYASHI
Affiliation:
Teaching and Research Support Center, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
Y. OKADA
Affiliation:
Department of Tropical Medicine and Parasitology, Keio University School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan
J. ITOH
Affiliation:
Department of Tropical Medicine and Parasitology, Keio University School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan
T. TAKEUCHI
Affiliation:
Teaching and Research Support Center, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan
*
*Corresponding author: Department of Infectious Diseases, Tokai University School of Medicine, Isehara, Kanagawa 259-1193, Japan. Tel: +81 463 93 1121. Fax: +81 463 95 5450. E-mail: [email protected]
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Summary

We have recently identified 2 surface proteins in Entamoeba histolytica as intermediate subunits of galactose- and N-acetyl-D-galactosamine-inhibitable lectin (EhIgl1 and EhIgl2); these proteins both contain multiple CXXC motifs. Here, we report the molecular characterization of the corresponding proteins in Entamoeba dispar, which is neither pathogenic nor invasive. Two Igl genes encoding 1110 and 1106 amino acids (EdIgl1 and EdIgl2) were cloned from 2 strains of E. dispar. The amino acid sequence identities were 79% between EdIgl1 and EdIgl2, 75–76% between EdIgl1 and EhIgl1, and 73–74% between EdIgl2 and EhIgl2. However, all the CXXC motifs were conserved in the EdIgl proteins, suggesting that the fold conferred by this motif is important for function. Comparison of the expression level of the Igl genes by real-time RT-PCR showed 3–5 times higher expression of EdIgl1 compared to EdIgl2. Most EdIgl1 and EdIgl2 proteins were co-localized on the surface and in the cytoplasm of trophozoites, based on confocal microscopy. However, a different localization of EdIgl1 and EdIgl2 in intracellular vacuoles and a different level of phenotypic expression of the two Igls were also observed. These results demonstrate that Igls are important proteins even in non-pathogenic amoeba and that Igl1 and Igl2 may possess different functions.

Keywords

Entamoeba disparEntamoeba histolyticaintermediate subunit of Gal/GalNAc lectin (Igl)cysteine-rich protein
Type
Research Article
Information
Parasitology , Volume 134 , Issue 14 , December 2007 , pp. 1989 - 1999
Copyright
Copyright © Cambridge University Press 2007

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References

REFERENCES

Bruchhaus, I., Jacobs, T., Leippe, M. and Tannich, E. (1996). Entamoeba histolytica and Entamoeba dispar: differences in numbers and expression of cysteine proteinase genes. Molecular Microbiology 22, 255–263.CrossRefGoogle ScholarPubMed
Cheng, X.-J., Hughes, M. A., Huston, C. D., Loftus, B., Gilchrist, C. A., Lockhart, L. A., Ghosh, S., Miller-Sims, V., Mann, B. J., Petri, W. A. Jr. and Tachibana, H. (2001). Intermediate subunit of the Gal/GalNAc lectin of Entamoeba histolytica is a member of a gene family containing multiple CXXC sequence motifs. Infection and Immunity 69, 5892–5898.CrossRefGoogle ScholarPubMed
Cheng, X.-J., Kaneda, Y. and Tachibana, H. (1997). A monoclonal antibody against the 150-kDa surface antigen of Entamoeba histolytica inhibits adherence and cytotoxicity to mammalian cells. Medical Science Research 25, 159–161.Google Scholar
Cheng, X.-J. and Tachibana, H. (2001). Protection of hamsters from amebic liver abscess formation by immunization with the 150- and 170-kDa surface antigens of Entamoeba histolytica. Parasitology Research 87, 126–130.CrossRefGoogle ScholarPubMed
Cheng, X.-J., Tachibana, H. and Kaneda, Y. (1999). Protection of hamsters from amebic liver abscess formation by a monoclonal antibody to a 150-kDa surface lectin of Entamoeba histolytica. Parasitology Research 85, 78–80.CrossRefGoogle ScholarPubMed
Cheng, X.-J., Tsukamoto, H., Kaneda, Y. and Tachibana, H. (1998). Identification of the 150-kDa surface antigen of Entamoeba histolytica as a galactose- and N-acetyl-D-galactosamine-inhibitable lectin. Parasitology Research 84, 632–639.Google Scholar
Davis, P. H., Schulze, J. and Stanley, S. L. Jr. (2007). Transcriptomic comparison of two Entamoeba histolytica strains with defined virulence phenotypes identifies new virulence factor candidates and key differences in the expression patterns of cysteine proteases, lectin light chains, and calmodulin. Molecular and Biochemical Parasitology 151, 118–128.Google Scholar
Diamond, L. S. and Clark, C. G. (1993). A redescription of Entamoeba histolytica Schaudinn, 1903 (Emended Walker, 1911) separating it from Entamoeba dispar Brumpt, 1925. Journal of Eukaryotic Microbiology 40, 340–344.CrossRefGoogle ScholarPubMed
Diamond, L. S., Harlow, D. R. and Cunnick, C. C. (1978). A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Transactions of the Royal Society of Tropical Medicine and Hygiene 72, 431–432.CrossRefGoogle ScholarPubMed
Hernández-Ramírez, V. I., Rios, A., Angel, A., Magos, M. A., Pérez-Castillo, L., Rosales-Encina, J. L., Castillo-Henkel, E. and Talamás-Rohana, P. (2007). Subcellular distribution of the Entamoeba histolytica 140 kDa FN-binding molecule during host-parasite interaction. Parasitology 134, 169–177.CrossRefGoogle ScholarPubMed
Khalifa, S. A., Imai, E., Kobayashi, S., Haghighi, A., Hayakawa, E. and Takeuchi, T. (2006). Growth-promoting effect on iron-sulfur proteins on axenic cultures of Entamoeba dispar. Parasite 13, 51–58.CrossRefGoogle ScholarPubMed
Kobayashi, S., Imai, E., Haghighi, A., Khalifa, S. A., Tachibana, H. and Takeuchi, T. (2005). Axenic cultivation of Entamoeba dispar in newly designed yeast extract-iron-gluconic acid-dihydroxyacetone-serum medium. Journal of Parasitology 91, 1–4.Google Scholar
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227, 680–685.Google Scholar
MacFarlane, R. C. and Singh, U. (2006). Identification of differentially expressed genes in virulent and nonvirulent Entamoeba species: potential implications for amebic pathogenesis. Infection and Immunity 74, 340–351.CrossRefGoogle ScholarPubMed
McCoy, J. J. and Mann, B. J. (2005). Proteomic analysis of Gal/GalNAc lectin-associated proteins in Entamoeba histolytica. Experimental Parasitology 110, 220–225.CrossRefGoogle ScholarPubMed
Mitra, B. N., Yasuda, T., Kobayashi, S., Saito-Nakano, Y. and Nozaki, T. (2005). Differences in morphology of phagosomes and kinetics of acidification and degradation in phagosomes between the pathogenic Entamoeba histolytica and the non-pathogenic Entamoeba dispar. Cell Motility and the Cytoskeleton 62, 84–99.CrossRefGoogle ScholarPubMed
Okada, M., Huston, C. D., Mann, B. J., Petri, W. A. Jr.Kita, K. and Nozaki, T. (2005). Proteomic analysis of phagocytosis in the enteric protozoan parasite Entamoeba histolytica. Eukaryotic Cell 4, 827–831.CrossRefGoogle ScholarPubMed
Okada, M., Huston, C. D., Oue, M., Mann, B. J., Petri, W. A. Jr.Kita, K. and Nozaki, T. (2006). Kinetics and strain variation of phagosome proteins of Entamoeba histolytica by proteomic analysis. Molecular and Biochemical Parasitology 145, 171–183.Google Scholar
Petri, W. A. Jr.Chapman, M. D., Snodgrass, T., Mann, B. J., Broman, J. and Ravdin, J. I. (1989). Subunit structure of the galactose and N-acetyl-D-galactosamine-inhibitable adherence lectin of Entamoeba histolytica. Journal of Biological Chemistry 264, 3007–3012.Google Scholar
Petri, W. A. Jr.Haque, R. and Mann, B. J. (2002). The bittersweet interface of parasite and host: lectin-carbohydrate interactions during human invasion by the parasite Entamoeba histolytica. Annual Review of Microbiology 56, 39–64.CrossRefGoogle ScholarPubMed
Pillai, D. R., Britten, D., Ackers, J. P., Ravdin, J. I. and Kain, K. C. (1997). A gene homologous to hgl2 of Entamoeba histolytica is present and expressed in Entamoeba dispar. Molecular and Biochemical Parasitology 87, 101–105.Google Scholar
Pillai, D. R., Kobayashi, S. and Kain, K. C. (2001). Entamoeba dispar: molecular characterization of the galactose/N-acetyl-D-galactosamine lectin. Experimental Parasitology 99, 226–234.CrossRefGoogle ScholarPubMed
Seigneur, M., Mounier, J., Prevost, M. C. and Guillen, N. (2005). A lysine- and glutamic acid-rich protein, KERP1, from Entamoeba histolytica binds to human enterocytes. Cellular Microbiology 7, 569–579.Google Scholar
Shah, P. H., MacFarlane, R. C., Bhattacharya, D., Matese, J. C., Demeter, J., Stroup, S. E. and Singh, U. (2005). Comparative genomic hybridizations of Entamoeba strains reveal unique genetic fingerprints that correlate with virulence. Eukaryotic Cell 4, 504–515.CrossRefGoogle ScholarPubMed
Tachibana, H., Cheng, X.-J., Masuda, G., Horiki, N. and Takeuchi, T. (2004). Evaluation of recombinant fragments of Entamoeba histolytica Gal/GalNAc lectin intermediate subunit for serodiagnosis of amebiasis. Journal of Clinical Microbiology 42, 1069–1074.CrossRefGoogle ScholarPubMed
Tachibana, H., Ihara, S., Kobayashi, S., Kaneda, Y., Takeuchi, T. and Watanabe, Y. (1991). Differences in genomic DNA sequences between pathogenic and nonpathogenic isolates of Entamoeba histolytica identified by polymerase chain reaction. Journal of Clinical Microbiology 29, 2234–2239.CrossRefGoogle ScholarPubMed
Tachibana, H., Kobayashi, S., Cheng, X.-J. and Hiwatashi, E. (1997). Differentiation of Entamoeba histolytica from E. dispar facilitated by monoclonal antibodies against a 150-kDa surface antigen. Parasitology Research 83, 435–439.CrossRefGoogle Scholar
Talamás-Rohana, P., Rosales-Encina, J. L., Gutierrez, M. C. and Hernández, V. I. (1992). Identification and partial purification of an Entamoeba histolytica membrane protein that binds fibronectin. Archives of Medical Research 23, 119–123.Google Scholar
Walsh, J. A. (1986). Problems in recognition and diagnosis of amebiasis: estimation of the global magnitude of morbidity and mortality. Reviews of Infectious Diseases 8, 228–238.CrossRefGoogle ScholarPubMed
Weber, C., Guigon, G., Bouchier, C., Frangeul, L., Moreira, S., Sismeiro, O., Gouyette, C., Mirelman, D., Coppee, J. Y. and Guillen, N. (2006). Stress by heat shock induces massive down regulation of genes and allows differential allelic expression of the Gal/GalNAc lectin in Entamoeba histolytica. Eukaryotic Cell 5, 871–875.CrossRefGoogle ScholarPubMed
Willhoeft, U., Hamann, L. and Tannich, E. (1999). A DNA sequence corresponding to the gene encoding cysteine proteinase 5 in Entamoeba histolytica is present and positionally conserved but highly degenerated in Entamoeba dispar. Infection and Immunity 67, 5925–5929.CrossRefGoogle Scholar