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

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 x 10(8) cells). ATP hydrolysis was stimulated by MgCl2, and the Mg-dependent ecto-ATPase activity was 27.15 +/- 2.91 nmol Pi/h x 10(8) 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 Km 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.

Secreted phosphatase activity induced by dimethyl sulfoxide in Herpetomonas samuelpessoai.

A phosphatase activity of the trypanosomatid parasite Herpetomonas samuelpessoai was characterized using intact living cells. The effects of dimethyl sulfoxide (DMSO) on this activity were investigated. This phosphatase activity (2.53+/-0.01 nmol P(i)/mg protein x min) was linear with cell density and with time for at least 60 min. The optimum pH for the H. samuelpessoai phosphatase lies in the acid range. This phosphatase activity was inhibited by metal chelators and classical phosphatase inhibitors. A robust stimulation of the phosphatase activity was observed when the flagellates were grown in the presence of 4% DMSO, both when intact flagellates and when culture supernatant from those cells were assayed, as observed by biochemical and cytochemical analysis. We also demonstrate that DMSO induced the secretion and/or shedding of this phosphatase to the extracellular medium, with a possible involvement of protein kinase C in this process.