Proteomic analysis of two Trypanosoma cruzi zymodeme 3 strains

Abstract

Two Trypanosoma cruzi Z3 strains, designated as 3663 and 4167, were previously isolated from insect vectors captured in the Brazilian Amazon region. These strains exhibited different infection patterns in Vero, C6/36, RAW 264.7 and HEp-2 cell lineages, in which 3663 trypomastigote form was much less infective than 4167 ones. A proteomic approach was applied to investigate the differences in the global patterns of protein expression in these two Z3 strains. Two-dimensional (2D) protein maps were generated and certain spots were identified by mass spectrometry (MS). Our analyses revealed a significant difference in the expression profile of different proteins between strains 3663 and 4167. Among them, cruzipain, an important regulator of infectivity. This data was corroborated by flow cytometry analysis using anti-cruzipain antibody. This difference could contribute to the infectivity profiles observed for each strain by in vitro assay using different cell lines.

Repertoire, Genealogy and Genomic Organization of Cruzipain and Homologous Genes in Trypanosoma cruzi, T. cruzi-Like and Other Trypanosome Species

“…Differences in cruzipain expression were correlated with differential virulence for mice of T. cruzi isolates of Z3 (TcIII and IV) [50]. A proteomic analysis suggested that significant differences in the expression of cruzipain by isolates of TcIII and TcIV (Z3) could also contribute to their differential infectivity to cells [51]. Interestingly, proteolytic activity was lower in the virulent T. cruzi CL strain compared to the non-virulent CL-14 clone [52].…”

Proteomic analysis of differentially expressed proteins in the marine fish parasitic ciliate Cryptocaryon irritans

“…For example, serine proteases, DNase II and trypsin were identified as the early specific diagnostic antigens of trichinellosis by twodimensional electrophoresis (2-DE) and Western blot . Different strains of Trypanosoma cruzi zymodeme were analyzed by 2-DE and cruzipain was identified as an important antigen among these proteins from T. cruzi (Kikuchi et al, 2010). In this study, we performed differential proteomics and immunoproteomics approaches to identify differentially expressed proteins in three developmental stages, and immunogenic proteins were recognized by rabbit and grouper anti-sera.…”

The Expected Outcome of the Trypanosoma cruzi Proteomic Map: A Review of Its Potential Biological Applications for Drug Target Discovery

“…This enzyme is involved in the conversion of L-arginine to phosphoarginine. It also participates in the oxidative response and infectivity, as it was upregulated in two highly virulent strains (Kikuchi et al 2010 ). The absence of arginine kinase in mammalian cells, together with its importance in parasite ATP synthesis, credits this enzyme as a potential target for new chemotherapies (Silber et al 2005 ;Sodré et al 2009 ).…”

“…In trypanosomatids, genomic analysis has demonstrated the presence of diverse calpains (Ersfeld et al 2005 ), whose function is associated with differentiation and infectivity in T. cruzi (Ennes-Vidal et al 2010. Evidence of the presence of a calpain-like cysteine peptidase in different parasite forms, together with the absence of a similar sequence in humans, suggests its potential as a drug target (Atwood et al 2005 ;Andrade et al 2008 ;Sant'Anna et al 2009 ;Sodré et al 2009 ;Kikuchi et al 2010 ).…”

“…A preliminary comparative assessment of the proteomic profi le was performed between epimastigotes of isolates 3663 ( T. cruzi III) and 4167 ( T. cruzi IV) and reference strains CL Brener and Dm28c by 2-DE and MALDI-TOF/TOF (Kikuchi et al 2010 ). Fifty-seven proteins identifi ed from these isolates were involved in different cellular functions, such as parasite metabolism, protein assembly, stress response and cell-to-cell communication.…”