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CrATP as a new inhibitor of ecto-ATPases of trypanosomatids

Published online by Cambridge University Press:  07 January 2009

O. C. MOREIRA ,
P. F. RIOS ,
F. F. ESTEVES ,
J. R. MEYER-FERNANDES  and
H. BARRABIN
O. C. MOREIRA
Affiliation:
Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de Janeiro, Brasil
P. F. RIOS
Affiliation:
Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de Janeiro, Brasil
F. F. ESTEVES
Affiliation:
Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de Janeiro, Brasil
J. R. MEYER-FERNANDES
Affiliation:
Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de Janeiro, Brasil
H. BARRABIN*
Affiliation:
Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Centro de Ciências da Saúde, Universidade Federal do Rio de JaneiroRio de Janeiro, Brasil
*
*Corresponding author: Instituto de Bioquímica Médica, CCS, Universidade Federal do Rio de Janeiro, Cidade Universitária, CEP 21941-590, Rio de Janeiro, RJ, Brazil. Fax: +55 21 2270 8647. E-mail: [email protected]
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Summary

Trypanosomatid protozoa include heteroxenic species some of them pathogenic for men, animals and plants. Parasite membrane contains ecto-enzymes whose active sites face the external medium rather than the cytoplasm. Herpetomonas sp. displayed a Mg2+-dependent ecto-ATPase activity, a Mg-independent ecto-ADPase and an ecto-phosphatase activity. Both, the ecto-ADPase and phosphatase activities were insensitive to CrATP (chromium(III) adenosine 5′-triphosphate complex). Ecto-ATPase activity was reversibly inhibited. At 2 mm ATP the apparent Ki was 4·7±1·0 μm but a fraction of about 40–50% was insensitive to CrATP. Remarkably, at low substrate concentration (0·2 mm) more than 90% of the ecto-ATPase was inhibited with Ki=0·33±0·10 μm. These parameter dependences are interpreted as the presence of 2 ecto-ATPases activities, one of them with high ATP apparent affinity and sensitivity to CrATP. DIDS (4,4 diisothiocyanatostilbene 2,2′ disulfonic acid), suramin and ADP were also effective as inhibitors. Only ADP presented no additive inhibition with CrATP. The pattern of partial inhibition by CrATP was also observed for the ecto-ATPase activities of Leishmania amazonensis, Trypanosoma cruzi and Trypanosoma rangeli. CrATP emerges as a new inhibitor of ecto-ATPases and as a tool for a better understanding of properties and role of ecto-ATPases in the biology of parasites.

Keywords

ecto-ATPaseHerpetomonasCrATPtrypanosomatidsecto-phosphatase
Type
Research Article
Information
Parasitology , Volume 136 , Issue 1 , January 2009 , pp. 35 - 44
Copyright
Copyright © 2009 Cambridge University Press

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