Resumen

VILLAMIL-SILVA, Sharon Eliana et al. Identification of a cytoplasmic tryparedoxin peroxidase from Leishmania braziliensis. Rev.Colomb.Quim. [online]. 2021, vol.50, n.2, pp.3-14.  Epub 09-Sep-2021. ISSN 0120-2804.  https://doi.org/10.15446/rev.colomb.quim.v50n2.91721.

The antioxidant defense systems used by the intracellular parasite Leishmania braziliensis during the infection process make it possible to eliminate reactive oxygen and nitrogen species at the expense of reducing equivalents derived from trypanothione, avoiding cellular damage of the pathogen. In order to identify potential molecular targets for the development of drugs against this parasite, the cytoplasmic tryparedoxin peroxidase of L. braziliensis (LbTXNPxII), which is essential to reduce toxic concentrations of hydrogen peroxide in the context of infection, was carried out. In this regard, polyclonal antibodies were generated in an avian model, starting from the cloning, expression, and purification of the recombinant protein 6xHis-SUMO-LbTXNPxII (37kDa) in the heterologous system of Escherichia coli. The purified protein was used as an antigen for the production of IgY antibodies, whose implementation in in situ experiments allowed the detection and localization of the endogenous LbTXNPxII enzyme (22kDa) in the cytoplasm of fixed promastigotes, as well as the verification of its molecular interaction with nicotinamide/nicotinate mononucleotide adenylyltransferase, an enzyme involved in the synthesis of NAD. Thus, the development of a biochemical tool for the identification and study of the LbTXNPxII enzyme and its participation in energy metabolism and antioxidant defense pathways is reported.

Palabras clave : IgY polyclonal antibodies; Leishmania braziliensis; protein-protein interaction; subcellular localization; Tryparedoxin peroxidase.

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