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Mol Biochem Parasitol. 2013 Aug;190(2):63-75. doi: 10.1016/j.molbiopara.2013.06.006. Epub 2013 Jul 4.

Comparative proteomic analysis of antimony-resistant and -susceptible Leishmania braziliensis and Leishmania infantum chagasi lines.

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1
Laboratório de Parasitologia Celular e Molecular, Centro de Pesquisas René Rachou CPqRR/Fiocruz, Belo Horizonte, MG, Brazil.

Abstract

The emergence of drug-resistant Leishmania species is a significant problem in several countries. A comparative proteomic analysis of antimony-susceptible and antimony-resistant Leishmania braziliensis (LbSbR) and Leishmania infantum chagasi (LcSbR) lines was carried out using two-dimensional gel electrophoresis (2-DE) followed by mass spectrometry (LC/MS/MS) for protein identification. Out of 132 protein spots exclusive or up-regulated submitted to MS, we identified 80 proteins that corresponded to 57 distinct proteins. Comparative analysis of data showed that most of the protein spots with differential abundance in both species are involved in antioxidant defense, general stress response, glucose and amino acid metabolism, and cytoskeleton organization. Five proteins were commonly more abundant in both SbIII-resistant Leishmania lines: tryparedoxin peroxidase, alpha-tubulin, HSP70, HSP83, and HSP60. Analysis of the protein abundance by Western blotting assays confirmed our proteomic data. These assays revealed that cyclophilin-A is less expressed in both LbSbR and LcSbR lines. On the other hand, the expression of pteridine reductase is higher in the LbSbR line, whereas tryparedoxin peroxidase is overexpressed in both LbSbR and LcSbR lines. Together, these results show that the mechanism of antimony-resistance in Leishmania spp. is complex and multifactorial.

KEYWORDS:

2-DE; Antimony resistance; Cyclophilin-A; L. (L.) infantum chagasi SbIII-resistant; L. (L.) infantum chagasi Wild-type; L. (V.) braziliensis SbIII-resistant; L. (V.) braziliensis Wild-type; LbSbR; LbWTS; LcSbR; LcWTS; Leishmania braziliensis; Leishmania infantum chagasi; MS; Proteome; Pteridine reductase; SbIII; Tryparedoxin peroxidase; mass spectrometry; potassium antimonyl tartrate; two-dimensional gel electrophoresis

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