Hostname: page-component-586b7cd67f-gb8f7 Total loading time: 0 Render date: 2024-11-24T10:37:31.824Z Has data issue: false hasContentIssue false
  • English
  • Français

Characterization and localization of Schistosoma mansoni calreticulin Sm58

Published online by Cambridge University Press:  06 April 2009

J. Khalife ,
J. L. Liu ,
R. Pierce ,
E. Porchet ,
C. Godin  and
A. Capron
J. Khalife
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
J. L. Liu
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
R. Pierce
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
E. Porchet
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
C. Godin
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
A. Capron
Affiliation:
Centre d'Immunologie et de Biologie Parasitaire, INSERM U 167-CNRS 624, Institut Pasteur, 1 Rue de Professeur Calmette, B.P. 245, 59019 Lille Cédex, France
Get access Rights & Permissions [Opens in a new window]

Summary

Recombinant Schistosoma mansoni calreticulin (SmCaR) was expressed in Escherichia coli, using the glutathione S-transferase fusion protein, and its Ca2+-binding capacity was determined. Results obtained by a 45Ca2+ overlay technique showed that Ca2+-binding site(s) were present in the recombinant CaR indicating that proper folding of the protein was obtained using this system. An antiserum raised against the recombinant SmCaR showed that the native protein (Sm58) was expressed in all stages of the life-cycle from cercariae to the adult worm and in the egg. However, SmCaR seems to be a developmentally regulated protein whose expression can be used to study the post-transformational differentiation of the schistosomulum. Localization of SmCaR demonstrated that the majority of SmCaR was expressed in the epithelia of the digestive duct and in the genital organs. These results suggest that SmCaR, by regulating the Ca2+ concentration, may play an important role during cell proliferation. Finally the presence of SmCaR in miracidia and in the genital organs suggests that the antibody response directed against this protein could interfere in egg production.

Keywords

Schistosoma mansonicalreticulinCa2+-binding protein
Type
Research Article
Information
Parasitology , Volume 108 , Issue 5 , June 1994 , pp. 527 - 532
Copyright
Copyright © Cambridge University Press 1994

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Baksh, S. & Michalak, M. (1991). Expression of calreticulin in Escherichia coli and identification of its Ca2+ binding domains. Journal of Biological Chemistry 266, 21458–65.CrossRefGoogle ScholarPubMed
Burgogne, R. D. & Geisow, M. J. (1989). The annexin family of calcium-binding proteins. Cell Calcium 10, 1–10.CrossRefGoogle Scholar
Burns, K., Helgason, C. D., Bleackley, R. C. & Michalak, M. (1992). Calreticulin in T lymphocytes: identification of calreticulin in T lymphocytes and demonstration that activation of T cells correlates with increased levels of calreticulin mRNA and protein. Journal of Biological Chemistry 267, 19039–42.CrossRefGoogle Scholar
Davies, T. W. (1983). Schistosoma mansoni: the structure and elemental composition of the pre-acetabular penetration gland cell secretion in pre-emergent cercariae. Parasitology 87, 55–60.CrossRefGoogle ScholarPubMed
Dingwall, C., Dilworth, S. M., Black, S. J., Kearsey, S. E., Cox, L. S. & Laskey, R. A. (1987). Nucleoplasmin cDNA sequence reveals polyglutamic acid tracts and a cluster of sequences homologous to putative nuclear localization signals. European Molecular Biological Organisation Journal 6, 69–74.CrossRefGoogle Scholar
Dupuis, M., Schaerer, E., Krause, K. H. & Tschopp, J. (1993). The calcium-binding protein Calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. Journal of Experimental Medicine 177, 1–7.CrossRefGoogle Scholar
Fuhrman, J. A. (1990). Calcium-binding proteins in schistosomes. Parasitology Today 6, 172–3.CrossRefGoogle ScholarPubMed
Havercroft, J. C., Huggins, M. C., Dunne, D. W. & Taylor, D. W. (1990). Characterization of Sm20, a 20 kilodalton calcium-binding protein of Schistosoma mansoni. Molecular and Biochemical Parasitology 38, 211–20.CrossRefGoogle ScholarPubMed
Havercroft, J. C., Smith, A. L. & Williams, R. H. (1991).Schistosoma mansoni: immunolocalization of the calcium binding protein Sm20. Parasite Immunology 13, 593–604.CrossRefGoogle ScholarPubMed
Heizmann, C. W. & Hunziker, W. (1991). Intracellular calcium-binding proteins: more sites than insights. Trends in Biochemical Sciences 16, 98–103.CrossRefGoogle ScholarPubMed
Khalife, J., Trottein, F., Schacht, A. M., Godin, C., Pierce, R. J. & Capron, A. (1993). Cloning of the gene encoding a Schistosoma mansoni antigen homologous to human Ro/SS-A autoantigen. Molecular and Biochemical Parasitology 57, 193–202.CrossRefGoogle ScholarPubMed
Khalife, J., Grzych, J. M., Pierce, R. J., Ameisen, J. C., Schacht, A. M., Gras-Masse, H., Tartar, A., Lecocq, J. P. & Capron, A. (1990). Immunological cross- reactivity between the human immunodeficiency virus type I virion infectivity factor and a 170 kDa surface antigen of Schistosoma mansoni. Journal of Experimental Medicine 172, 1001–4.CrossRefGoogle Scholar
Klee, C. B. (1988). Ca2+-dependent phospholipid-(and membrane-) binding proteins Biochemistry 27, 6645–53.CrossRefGoogle ScholarPubMed
Kretsinger, R. H., Muncreif, N. D., Goodman, M. & Czelusniak, T. (1988). In The Calcium Channel, Structure, Function and Implication (ed. Morad, M., Naylor, W. G., Kazda, S. & Schramm, M.), pp. 16–34. New York: Springer Verlag.CrossRefGoogle Scholar
Maruyama, K., Mikawa, T. & Ebashi, S. (1984). Detection of calcium-binding proteins by 45Ca autoradiography on nitrocellulose membranes after sodium dodecyl sulfate gel electrophoresis. Journal of Biochemistry 95, 511–19.CrossRefGoogle ScholarPubMed
Michalak, M., Milner, R. E., Burns, K. & Opas, N. (1992). Calreticulin. The Biochemical Journal 285, 681–92.CrossRefGoogle ScholarPubMed
Milner, R. E., Baksh, S., Shemenko, C., Carpenter, R. M., Smilie, L., Vance, J. E., Opas, M. & Michalak, M. (1991). Calreticulin, and not calsequestrin, is the major calcium-binding protein of smooth muscle sarcoplasmic reticulin and liver endoplasmic reticulum. Journal of Biological Chemistry 266, 7155–65.CrossRefGoogle ScholarPubMed
Pelham, H. H., Hardwick, K. G. & Lewis, M. J. (1988). Sorting of soluble proteins in yeast. European Molecular Biological Organisation Journal 7, 1757–62.CrossRefGoogle ScholarPubMed
Siddiqui, A. A., Podestra, R. B. & Clarke, M. W. (1991). Schistosoma mansoni: characterization and identification of calcium-binding proteins associated with the apical plasma membrane and envelope. Experimental Parasitology 72, 63–8.CrossRefGoogle ScholarPubMed
Vaitukaitis, J., Robbins, J. B., Nieschlaf, E. & Ross, G. T. (1971). A method for producing specific antisera with small doses of immunogen. Journal of Clinical Endocrinology 17, 988–91.CrossRefGoogle Scholar