Proteomic analysis of metacyclic trypomastigotes undergoing Trypanosoma cruzi metacyclogenesis

Abstract

Trypanosoma cruzi, the causative agent of the Chagas disease, has a complex life cycle alternating between replicative and noninfective forms with nonreplicative and infective forms of the parasite. Metacyclogenesis is a process that takes place in the invertebrate host, comprising morphogenetic transformation from a noninfective form to an infective form, such that parasites acquire the ability to invade human cells. We analyze here the metacyclogenesis process by 2D electrophoresis coupled to MALDI-TOF MS. A large proportion of unique proteins expressed during metacyclogenesis were observed. Interestingly, 50% of the spots were found to differ between epimastigotes and trypomastigotes. We provide a 2D map of the infective metacyclic trypomastigotes. Sixty six protein spots were successfully identified corresponding to 43 different proteins. We analyzed the expression profiles for the identified proteins along metacyclogenesis and classified them into three groups according to their maximal level of expression. We detected several isoforms for a number of proteins, some displaying differential expression during metacyclogenesis. These results suggest that posttranslational modifications may be a fundamental part of the parasite's strategy for regulating gene expression during differentiation. This study contributes to the identification of relevant proteins involved in the metacyclogenesis process. The identification and molecular characterization of these proteins will render vital information about the steps of the parasite differentiation into the infective form.

Pathogenesis of Chagas' Disease: Parasite Persistence and Autoimmunity

“…Biochemical and molecular evidence confirmed the presence of basic proteins in the kinetoplast, and histone H1 participates in kDNA condensation (307,370). Fluorescence microscopy revealed bromodeoxyuridine (BrdU)-labeled free gaped minicircles (343), which replicate in approximate synchrony with the nuclear S phase.…”

Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis

“…However, metacyclogenesis can be mimicked in vitro using well-defined media that stimulate the differentiation process by simulating the vector's gut physicochemical conditions (10). This process results in important changes that have been studied intensively through proteomic approaches (11,12).…”

TcI Isolates of Trypanosoma cruzi Exploit the Antioxidant Network for Enhanced Intracellular Survival in Macrophages and Virulence in Mice

“…Likewise, members of the paraflagellar rod (PFR) and paraxonemal protein families, known to be essential for flagellar wave frequency and amplitude and forward motility of the parasite (reviewed in reference 44), were enhanced in abundance in COL and SYL (versus TCC) strains. Further, SYL and COL trypomastigotes also exhibited an increased abundance of calpain-like cysteine peptidases, surface glycoprotein gp82, and cruzipain that are involved in host cell attachment and invasion (45,46). We surmise that virulence of the COL, SYL, and possibly other isolates of the TcI DTU is determined by the motility capacity that ensures parasite transmission from triatomines and dissemination to tissues in the mammalian host.…”

“…The question arises as to how the TXNPx/TXN antioxidant network is associated with the virulence of the TcI isolates of T. cruzi. One explanation could be that, upon infection, T. cruzi faces the hostile environment of phagocytes, and an increase in antioxidant response would allow adaptation of the infective forms to the oxi- dative environment of phagocytic cells in the mammalian host (45,46). Activated macrophages provide a key defense response to the invading pathogens and exert cytotoxic effects via NADPH oxidase (NOX)-mediated production of superoxide and ROS that result in enhanced oxidative modification of DNA, proteins, and lipids (53,54).…”