TY - JOUR
T1 - Platelet-activating factor induction of secreted phosphatase activity in Trypanosoma cruzi
AU - Rodrigues, C. O.
AU - Dutra, P. M.L.
AU - Barros, F. S.
AU - Souto-Padrón, T.
AU - Meyer-Fernandes, J. R.
AU - Lopes, A. H.C.S.
N1 - Funding Information:
We thank Dr. Scott M. Landfear for critical comments on the manuscript; Dr. Renato S. B. Cordeiro, Dr. Mario A. C. Silva-Neto, and Dr. Norton Heise for helpful discussions; and Mr. Luciano A. M. Grillo and Mr. Felipe A. Dias for technical assistance. This work was supported by the Brazilian agencies PRONEX (0885), FAPERJ, FINEP and CNPq.
PY - 1999/12/9
Y1 - 1999/12/9
N2 - The effects of platelet-activating factor (PAF) on the ecto-phosphatase activity of Trypanosoma cruzi were investigated. Living parasites hydrolyzed p-nitrophenylphosphate (p-NPP) at a rate of 5.71 ± 0.37 nmol P(i) mg-1 min-1. This ecto-phosphatase activity increased to 8.70 ± 1.12 nmol P(i) mg-1 min-1 when the cells were grown in the presence of 10-9 M PAF. This effect was probably due to stimulation of the release of the ecto-phosphatase and/or the secretion of an intracellular phosphatase to the extracellular medium, as suggested by cytochemical analysis. Modulation of the ecto-phosphatase activity was also observed when PAF was added during the time course of the reaction. WEB 2086, a competitive PAF antagonist, was able to revert PAF effects when both were used at the same concentration. When PAF was added to a membrane enriched fraction preparation of T. cruzi, no alteration on the phosphatase activity was observed. This result suggests an involvement of intracellular signaling, as PAF was only effective on intact cells. Sphingosine and phorbol-12-myristate-13-acetate (PMA) were then used to investigate a possible involvement of protein kinase C (PKC) with PAF-induced phosphatase secretion. Sphingosine by itself stimulated the secretion of a phosphatase but did not significantly interfere with PAF effects on this enzyme. On the other hand, PMA was able to abrogate PAF-induced release of this phosphatase. These data are highly suggestive of a putative involvement of signal transduction mediated by a ligand of mammalian origin (PAF), through PKC and a specific receptor located on the cell surface of the human parasite Trypanosoma cruzi.
AB - The effects of platelet-activating factor (PAF) on the ecto-phosphatase activity of Trypanosoma cruzi were investigated. Living parasites hydrolyzed p-nitrophenylphosphate (p-NPP) at a rate of 5.71 ± 0.37 nmol P(i) mg-1 min-1. This ecto-phosphatase activity increased to 8.70 ± 1.12 nmol P(i) mg-1 min-1 when the cells were grown in the presence of 10-9 M PAF. This effect was probably due to stimulation of the release of the ecto-phosphatase and/or the secretion of an intracellular phosphatase to the extracellular medium, as suggested by cytochemical analysis. Modulation of the ecto-phosphatase activity was also observed when PAF was added during the time course of the reaction. WEB 2086, a competitive PAF antagonist, was able to revert PAF effects when both were used at the same concentration. When PAF was added to a membrane enriched fraction preparation of T. cruzi, no alteration on the phosphatase activity was observed. This result suggests an involvement of intracellular signaling, as PAF was only effective on intact cells. Sphingosine and phorbol-12-myristate-13-acetate (PMA) were then used to investigate a possible involvement of protein kinase C (PKC) with PAF-induced phosphatase secretion. Sphingosine by itself stimulated the secretion of a phosphatase but did not significantly interfere with PAF effects on this enzyme. On the other hand, PMA was able to abrogate PAF-induced release of this phosphatase. These data are highly suggestive of a putative involvement of signal transduction mediated by a ligand of mammalian origin (PAF), through PKC and a specific receptor located on the cell surface of the human parasite Trypanosoma cruzi.
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U2 - 10.1006/bbrc.1999.1759
DO - 10.1006/bbrc.1999.1759
M3 - Article
C2 - 10581161
AN - SCOPUS:0033539896
VL - 266
SP - 36
EP - 42
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -