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Genome-Wide Proteomics and Phosphoproteomics Analysis of Trypanosoma cruzi During Differentiation

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Trypanosomatids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2116))

Abstract

Trypanosoma cruzi is a pathogenic protozoan that still has an impact on public health, despite the decrease in the number of infection cases along the years. T. cruzi possesses an heteroxenic life cycle in which it differentiates in at least four forms. Among the differentiation processes, metacyclogenesis has been exploited in different views by researchers. An intriguing question that rises is how metacyclogenesis is triggered and controlled by cell signaling and which are the differentially expressed proteins and posttranslational modifications involved in this process. An important cell signaling pathway is the protein phosphorylation, and it is reinforced in T. cruzi in which the gene expression control occurs almost exclusively posttranscriptionally. Additionally, the number of protein kinases in T. cruzi is relatively high compared to other organisms. A way to approach these questions is evaluating the cells through phosphoproteomics and proteomics. In this chapter, we will describe the steps from the cell protein extraction, digestion and fractionation, phosphopeptide enrichment, to LC-MS/MS analysis as well as a brief overview on peptide identification. In addition, a published method for in vitro metacyclogenesis will be detailed.

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Acknowledgments

The authors are very grateful to Dr. Rafael Luis Kessler for kindly providing the microscopy images of T. cruzi and to MSc Wagner Nagib de Souza Birbeire for assistance in artwork design.

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Authors and Affiliations

  1. Carlos Chagas Institute, Fiocruz Parana, Curitiba, Parana, Brazil

    Michel Batista, Aline Castro Rodrigues Lucena & Fabricio Klerynton Marchini

  2. Faculty of Medicine, Catholic University of Cuenca, Cuenca, Azuay, Ecuador

    Juliana Carolina Amorim

  3. Department of Cancer Biology, Metastasis Research Center, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Fernanda Grande Kugeratski

  4. Molecular Biology Institute of Parana, Curitiba, Parana, Brazil

    Carla Vanessa de Paula Lima

Authors
  1. Michel Batista
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  2. Juliana Carolina Amorim
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  3. Aline Castro Rodrigues Lucena
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  4. Fernanda Grande Kugeratski
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  5. Carla Vanessa de Paula Lima
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  6. Fabricio Klerynton Marchini
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Corresponding author

Correspondence to Fabricio Klerynton Marchini .

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Editors and Affiliations

  1. School of Biological Sciences, University of Edinburgh, Edinburgh, UK

    Paul A. M. Michels

  2. School of Applied Sciences, University of Huddersfield, Huddersfield, UK

    Michael L. Ginger

  3. Faculty of Biology, Technion–Israel Institute of Technology, Haifa, Israel

    Dan Zilberstein

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Batista, M., Amorim, J.C., Lucena, A.C.R., Kugeratski, F.G., de Paula Lima, C.V., Marchini, F.K. (2020). Genome-Wide Proteomics and Phosphoproteomics Analysis of Trypanosoma cruzi During Differentiation. In: Michels, P., Ginger, M., Zilberstein, D. (eds) Trypanosomatids. Methods in Molecular Biology, vol 2116. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0294-2_11

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  • DOI: https://doi.org/10.1007/978-1-0716-0294-2_11

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