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Purification, characterization, and immunolocalization of paramyosin from the adult stage of Fasciola hepatica

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Abstract

Paramyosin, a vaccine candidate in different helminthiases, was purified from the adult liver fluke Fasciola hepatica using two different procedures. The first started with a crude extraction of paramyosin in high-salt buffer followed by gel filtration chromatography and two precipitation-solubilization cycles; in the second, anion exchange chromatography replaced the gel filtration step. In both cases, the apparent molecular weight of the purified protein determined by sodium dodecyl sulfate gel electrophoresis under reducing and non-reducing conditions was 97 kDa and 200 kDa, respectively. The molecular weights were consistent with the presence of a dimeric protein linked by disulfide bridges. Western blot analysis showed that the dimeric and monomeric forms were both recognized by an antiserum raised against the F. hepatica 97 kDa band (α-FhPmy), and by an anti-Schistosoma mansoni paramyosin immune serum. Immunohistochemistry using α-FhPmy demonstrated the localization of paramyosin within the subtegumental muscle and in muscle cells surrounding the gut of adult parasites. We also observed labeling of extramuscular structures like testes, surface lamellae of the gut and the tegument of adult flukes.

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Acknowledgements

We thank Dr. Edward Pearce from Cornell University, USA for kindly supplying the anti-SmPmy hyper-immune serum. This work was supported by grants from IFS (International Foundation for Science) and SIDA (Swedish International Development Agency). Patricia Berasain was the recipient of a PEDECIBA doctoral fellowship. The experiments described in this study comply with the current laws of Uruguay.

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Authors and Affiliations

  1. Dpto. de Biología Celular y Molecular, Facultad de Ciencias, Instituto de Higiene, Unidad de Biología Parasitaria, Av. A. Navarro 3051, C.P. 11600, Montevideo, Uruguay

    Martín Cancela, Carlos Carmona & Patricia Berasain

  2. Laboratorio de Inmunotecnología, Cátedra de Inmunología, Facultades de Química y Ciencias, Instituto de Higiene, Montevideo, Uruguay

    Silvina Rossi

  3. Department of Pathology and Psychiatry, New York University, New York, USA

    Blas Frangione & Fernando Goñi

  4. Cátedra de Inmunología, Facultad de Química, Universidad de la República Oriental del Uruguay, Montevideo, Uruguay

    Fernando Goñi

Authors
  1. Martín Cancela
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  2. Carlos Carmona
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  3. Silvina Rossi
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  4. Blas Frangione
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  5. Fernando Goñi
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  6. Patricia Berasain
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Correspondence to Patricia Berasain.

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Cancela, M., Carmona, C., Rossi, S. et al. Purification, characterization, and immunolocalization of paramyosin from the adult stage of Fasciola hepatica . Parasitol Res 92, 441–448 (2004). https://doi.org/10.1007/s00436-003-1059-3

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  • DOI: https://doi.org/10.1007/s00436-003-1059-3

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