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Comparative Two-Dimensional Gel Electrophoresis of Trypanosoma cruzi Mammalian-Stage Forms in an Alkaline pH Range

[ Vol. 22 , Issue. 12 ]

Author(s):

Adriana D. Magalhães, Rayner Myr L. Queiroz, Izabela M. D. Bastos, Jaime M. Santana, Marcelo V. Sousa, Carlos André O. Ricart and Sébastien Charneau   Pages 1066 - 1075 ( 10 )

Abstract:


It is estimated that several million people are currently infected worldwide by the protozoan parasite, Trypanosoma cruzi, which causes Chagas disease. After mammalian host infection, a fundamental event is the differentiation from infective trypomastigotes into replicative amastigotes (amastigogenesis) inside host-cells. To unravel the particularities of both forms, it is essential to identify molecules presented in each form. Since T. cruzi gene expression regulation occurs mainly at posttranscriptional level, a proteomic approach is appropriate. Due to intrinsic difficulties with performing 2-DE in the alkaline pH range, there are no reports on 2-DE-based comparative proteome analysis of T. cruzi mammalianstage forms that focus on alkaline polypeptides. Here, we performed a comparative proteome analysis between tissue culture- derived trypomastigotes and extracellular amastigote-like cells using conditions optimized for the 6-11 pH range followed by identification by MALDI-TOF/TOF technology. The alkaline 2-DE maps from both forms show that proteins with a pI above 7.0 were not underrepresented (= 65% of proteins detected). Moreover the differences in protein expression between the Human-hosted T. cruzi forms corroborated previous proteomic studies and corresponded to their biological traits.

Keywords:

Alkaline 2-DE, amastigote, Chagas disease, proteome, Trypanosoma cruzi, trypomastigote.

Affiliation:

Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasilia, Brasilia-DF, 70910-900, Brazil.

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