Aspartate is one of the compounds that induce the differentiation process of the non-infective epimastigote stage to the infective trypomastigote stage of the protozoan parasite Trypanosoma cruzi. l-aspartate is transported by both epimastigote and trypomastigote cells at the same rate, about 3.4 pmolmin(-1) per 10(7) cells. Aspartate transport is only competed by glutamate suggesting that this transport system is specific for anionic amino acids. Aspartate uptake rates increase along the parasite growth curve, by amino acids starvation or pH decrease. The metabolic fate of the transported aspartate was predicted in silico by identification of seven putative genes coding for enzymes involved in aspartate metabolism that could be related to the differentiation process.
L-Cysteine and methionine are unique amino acids that act as sulfur donors in all organisms. In the specific case of Trypanosomatids, L-cysteine is particularly relevant as a substrate in the synthesis of trypanothione. Although it can be synthesized de novo, L-cysteine is actively transported in Trypanosoma cruzi epimastigote cells. L-Cysteine uptake is highly specific; none of the amino acids assayed yield significant differences in terms of transport rates. L-Cysteine is transported by epimastigote cells with a calculated apparent K(m) of 49.5 microM and a V(max) of about 13 pmol min(-1) per 10(7) cells. This transport is finely regulated by amino acid starvation, extracellular pH, and between the parasite growth phases. In addition, L-cysteine is incorporated post-translationally into proteins, suggesting its role in iron-sulfur core formation. Finally, the metabolic fates of Lcysteine were predicted in silico.
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