The nutritional differences among the habitats where these parasites thrive lead to remarkable variations in their energy metabolism (Coustou et al., 2008; Tielens & van Hellemond, 2009). To survive, these pathogens depend on a complex network of low-molecular-mass oxidoreductases; the detoxification of reactive oxygen and nitrogen species is accomplished by a complex redox cascade which utilizes the reducing equivalents derived from trypanothione. The latter dithiol molecule is notably abundant in

these pathogens (in the mM range) and is maintained in its reduced form by trypanothione reductase, NADPH being the primary source of reducing power for these processes (for review see Irigoin et al., 2008; Krauth-Siegel see more & Comini, 2008). In these parasites, large amounts of NADPH are required for de novo synthesis of fatty acids (for review see Lee et al., 2007). These molecules are needed to build the glycosylphosphatidylinositol anchors which attach the KU-57788 mw large amounts of glycoconjugates that coat the surface of these parasites (Ferguson, 1997; Donelson, 2003). These pathogens have redundant pathways to maintain the required reducing power, and the pentose phosphate pathway is a potential target for drug design against trypanosomes

(Hanau et al., 2004; Igoillo-Esteve et al., 2007). Malic enzymes (MEs), putative isocitrate dehydrogenases and glutamate dehydrogenases are among the other NADP-linked enzymes that are good candidates to contribute to NADPH production. Early findings showed that T. cruzi contains two MEs, a cytosolic and a mitochondrial isoform. Although these enzymes have not been completely purified, the enriched fractions exhibited high affinities for NADP and the cytosolic isozyme were strongly activated by l-aspartate (Cannata

et al., 1979; Cazzulo et al., 1980). Similarly, in T. brucei two putative MEs are predicted to be functional; recent RNAi studies showed that at least one of these isozymes is essential for parasite survival (Coustou et al., 2008). However, none of the T. brucei MEs has been functionally characterized, although the activity of one isozyme was determined Cediranib (AZD2171) in procyclics (Opperdoes & Cottem, 1982). The results presented herein provide a comparative description of the biochemical properties of T. cruzi and T. brucei MEs. Although the MEs from both parasites exhibit the same subcellular localization, they differ in their kinetic properties and developmental expression patterns. Procyclic forms of T. brucei stock 427 were grown as described previously (Brun & Schonenberger, 1979). The bloodstream forms of T. brucei stock 427 were grown in male Wistar rats; trypanosomes were obtained by cardiac puncture and separated from blood constituents by DEAE-cellulose chromatography (Lanham & Godfrey, 1970). Trypanosoma cruzi (CL Brener clone) insect and mammalian stages were grown as previously described (Cazzulo et al., 1985; Franke de Cazzulo et al., 1994). Total DNA was isolated from T.