, 2006); it is assumed that the malate generated inside any Linsitinib of these organelles can be transported into the cytosol. Opposed to T. brucei, the insect stage of T. cruzi can only produce malate inside the glycosome and the mitochondrion, as in this parasite the cytosolic malate dehydrogenase (MDH) is replaced by an aromatic 2-hydroxyacid dehydrogenase that is unable to catalyze the reduction of oxaloacetate into malate (Cazzulo Franke et al., 1999). Besides, the expression of the glycosomal and mitochondrial MDHs is developmentally regulated in the American trypanosome, the glycosomal isozyme being downregulated in T. cruzi amastigotes. By contrast, the mitochondrial MDH (mMDH)

is expressed throughout the whole life cycle of parasite, and at the protein level the enzyme seems to be more abundant in amastigotes. Also, the mitochondrial aspartate aminotransferase is expressed in all T. cruzi stages, but appeared to be more

abundant in the intracellular amastigotes (Marciano et al., 2008). Although in T. cruzi amastigotes, l-aspartate is transaminated into oxaloacetate in the mitochondrion and the cytosol, the latter is converted into malate only through the action of mMDH. As the malate produced in the mitochondrion can be easily exported into the cytosol, this metabolite can serve as a substrate for both the mitochondrial and the cytosolic MEs. In summary, in T. cruzi the cytosolic level of malate is determined by the metabolic activity within Olaparib concentration the mitochondrion. This hypothesis fits in well with early reports that postulated that in T. cruzi,

amino acids are actively metabolized in the mitochondrion, generating the precursors needed for energy production as well as the intermediates required for metabolic processes that occur in the cytosol (Tielens & Van Hellemond, 1998). This work was performed with grants from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad de Buenos Aires (UBA, B-094) and Agencia Nacional de Promoción Científica y Tecnológica (Argentina). C.N. and J.J.C. are members of the Mirabegron Research Career from CONICET, A.E.L. is supported by a fellowship from CONICET and D.A.M. by a fellowship from the Universidad Nacional de General San Martin. Fig. S1. Sequence alignment of MEs from Trypanosoma cruzi and Trypanosoma brucei with selected orthologues from higher eukaryotes linked to NAD and NADP coenzymes. Putative malic enzymes from trypanosomes were aligned with Homo sapiens mitochondrial NAD-dependent malic enzyme (MAOM_HUMAN, Genebank accession number P23368) and pigeon cytosolic NADP-dependent malic enzyme (MAOX_COLLI, Genebank accession number P40927) using ClustalX default settings. T. brucei putative MEs (TbME1, Gene ID Tb11.02.3130 and TbME2, Gene ID Tb11.02.3120), T. cruzi putative MEs (TcME1a, Gene ID Tc00.1047053505183.20, TcME1b, Gene ID Tc00.1047053508647.270, TcME2a, Gene ID Tc00.