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Subcellular localization of an extracellular serine protease in Leishmania (Leishmania) amazonensis

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Abstract

Extracellular proteolytic activity was detected in a Leishmania (L.) amazonensis culture supernatant and a 56-kDa protein was purified using (NH4)2SO4 precipitation followed by affinity chromatography on aprotinin–agarose. A rabbit serum obtained against the 56-kDa extracellular serine protease was used in order to analyze its location in L. (L.) amazonensis parasites. Immunocytochemistry studies revealed that the enzyme is mainly found in the flagellar pocket and cytoplasmic vesicles of promastigote forms, whereas in amastigotes, it is located in electron-dense structures resembling megasomes. These results indicate that the 56-kDa serine protease is released into the extracellular environment through the flagellar pocket; and its intracellular location suggests either a correlated enzymatic activity or intracellular trafficking.

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Acknowledgements

This work was supported by grants from the Fundação de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ), the Fundação Oswaldo Cruz (FIOCRUZ, PAPES) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). We are grateful indebted to Mr. G. Alves for technical assistance on Leishmania cultures.

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Authors and Affiliations

  1. Laboratório de Bioquímica de Proteínas e Peptídeos, Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil

    R. E. Silva-Lopez & S. Giovanni-De-Simone

  2. Divisão de Biologia Celular, Pesquisa Básica, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brazil

    J. A. Morgado-Díaz

  3. Laboratório de Biologia Molecular e Doenças Endêmicas, Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil

    C. R. Alves

  4. Departamento de Ultra-estrutura e Biologia Celular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil

    S. Côrte-Real

  5. Departamento de Biologia Celular e Molecular, Instituto de Biologia, Universidade Federal Fluminense, Niterói, RJ, Brazil

    S. Giovanni-De-Simone

Authors
  1. R. E. Silva-Lopez
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  2. J. A. Morgado-Díaz
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  3. C. R. Alves
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  4. S. Côrte-Real
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  5. S. Giovanni-De-Simone
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Correspondence to R. E. Silva-Lopez.

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Silva-Lopez, R.E., Morgado-Díaz, J.A., Alves, C.R. et al. Subcellular localization of an extracellular serine protease in Leishmania (Leishmania) amazonensis . Parasitol Res 93, 328–331 (2004). https://doi.org/10.1007/s00436-004-1144-2

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  • DOI: https://doi.org/10.1007/s00436-004-1144-2

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