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Parasitology Research

, Volume 93, Issue 1, pp 41–50 | Cite as

A Mg-dependent ecto-ATPase is increased in the infective stages of Trypanosoma cruzi

  • José Roberto Meyer-FernandesEmail author
  • Jorge Saad-Nehme
  • Carlos E. Peres-Sampaio
  • Rodrigo Belmont-Firpo
  • Danielle F. R. Bisaggio
  • Luciana C. do Couto
  • André Luíz Fonseca de Souza
  • Angela H. S. C. Lopes
  • Thais Souto-Padrón
Original Paper

Abstract

In this work, we describe the ability of living epimastigotes of Trypanosoma cruzi to hydrolyze extracellular ATP. In these intact parasites, there was a low level of ATP hydrolysis in the absence of any divalent metal (2.42±0.31 nmol Pi/h×108 cells). ATP hydrolysis was stimulated by MgCl2, and the Mg-dependent ecto-ATPase activity was 27.15±2.91 nmol Pi/h×108 cells. The addition of MgCl2 to the extracellular medium increased the ecto-ATPase activity in a dose-dependent manner. This stimulatory activity was also observed when MgCl2 was replaced by MnCl2, but not by CaCl2 or SrCl2. The apparent K m for Mg-ATP2− was 0.61 mM, and free Mg2+ did not increase the ecto-ATPase activity. This ecto-ATPase activity was insensitive to the inhibitors of other ATPase and phosphatase activities. To confirm that this Mg-dependent ATPase was an ecto-ATPase, we used an impermeant inhibitor, DIDS (4, 4′.diisothiocyanostylbene 2′-2′-disulfonic acid) as well as suramin, an antagonist of P2 purinoreceptors and inhibitor of some ecto-ATPases. These two reagents inhibited the Mg2+-dependent ATPase activity in a dose-dependent manner. A comparison among the Mg2+-ecto-ATPase activities of the three forms of T. cruzi showed that the noninfective epimastigotes were less efficient at hydrolyzing ATP than the infective trypomastigote and amastigote stages.

Keywords

Trypanosoma cruzi Ecto-ATPase Virulence 

Notes

Acknowledgements

We would like to acknowledge the excellent technical assistance of Fabiano Ferreira Esteves. This work was partially supported by grants from the Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Financiadora de Estudos e Projetos (FINEP), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Programa de Núcleos de Excelência (PRONEX, grant 0885) and FUJB/UFRJ.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • José Roberto Meyer-Fernandes
    • 1
    Email author
  • Jorge Saad-Nehme
    • 1
  • Carlos E. Peres-Sampaio
    • 1
  • Rodrigo Belmont-Firpo
    • 1
  • Danielle F. R. Bisaggio
    • 2
  • Luciana C. do Couto
    • 2
  • André Luíz Fonseca de Souza
    • 1
  • Angela H. S. C. Lopes
    • 2
  • Thais Souto-Padrón
    • 2
  1. 1.Departamento de Bioquímica Médica, Instituto de Ciências BiomédicasUniversidade Federal do Rio de JaneiroRio de Janeiro Brazil
  2. 2.Instituto de Microbiologia Prof. Paulo de Góes CCSUniversidade Federal do Rio de Janeiro Rio de JaneiroBrazil

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