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Updated: Jun 21, 2026
Ultrastructural Expansion Microscopy in Three In Vitro Life Cycle Stages of Trypanosoma cruzi
Published on: May 12, 2023
Phosphoproteomic analysis of the human pathogen Trypanosoma cruzi at the epimastigote stage
Ernesto S Nakayasu1, Matthew R Gaynor, Tiago J P Sobreira
1The Border Biomedical Research Center, Department of Biological Sciences, University of Texas at El Paso, El Paso, TX 79968, USA.
Abstract:
Trypanosoma cruzi is the etiologic agent of Chagas disease, which affects millions of people in Latin America and has become a public health concern in the United States and areas of Europe. The possibility that kinase inhibitors represent novel anti-parasitic agents is currently being explored. However, fundamental understanding of the cell-signaling networks requires the detailed analysis of the involved phosphorylated proteins. Here, we have performed a comprehensive MS-based phosphorylation mapping of phosphoproteins from T. cruzi epimastigote forms. Our LC-MS/MS, dual-stage fragmentation, and multistage activation analysis has identified 237 phosphopeptides from 119 distinct proteins. Furthermore, 220 phosphorylation sites were unambiguously mapped: 148 on serine, 57 on threonine, and 8 on tyrosine. In addition, immunoprecipitation and Western blotting analysis confirmed the presence of at least seven tyrosine-phosphorylated proteins in T. cruzi. The identified phosphoproteins were subjected to Gene Ontology, InterPro, and BLAST analysis, and categorized based on their role in cell structure, motility, transportation, metabolism, pathogenesis, DNA/RNA/protein turnover, and signaling. Taken together, our phosphoproteomic data provide new insights into the molecular mechanisms governed by protein kinases and phosphatases in T. cruzi. We discuss the potential roles of the identified phosphoproteins in parasite physiology and drug development.
Insights
This study maps protein phosphorylation in Trypanosoma cruzi, identifying key phosphoproteins involved in parasite function. These findings offer insights into cell signaling and potential drug targets for Chagas disease.
Area of Science:
- Parasitology and molecular biology
- Proteomics and post-translational modifications
Background:
- Chagas disease, caused by Trypanosoma cruzi, is a significant public health issue.
- Understanding Trypanosoma cruzi cell signaling is crucial for developing novel anti-parasitic agents, particularly kinase inhibitors.
Purpose of the Study:
- To perform a comprehensive MS-based phosphorylation mapping of phosphoproteins in Trypanosoma cruzi epimastigote forms.
- To identify and characterize phosphoproteins involved in parasite physiology and signaling pathways.
Main Methods:
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) with dual-stage fragmentation and multistage activation.
- Immunoprecipitation and Western blotting to confirm tyrosine-phosphorylated proteins.
- Bioinformatic analysis including Gene Ontology, InterPro, and BLAST.
Main Results:
- Identified 237 phosphopeptides from 119 distinct proteins.
- Mapped 220 phosphorylation sites (148 serine, 57 threonine, 8 tyrosine).
- Confirmed at least seven tyrosine-phosphorylated proteins and categorized identified proteins by function.
Conclusions:
- The phosphoproteomic data provide novel insights into Trypanosoma cruzi molecular mechanisms regulated by protein kinases and phosphatases.
- Identified phosphoproteins have potential roles in parasite physiology, pathogenesis, and drug development targeting Chagas disease.
