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Fluorescence Proteomic Technology to Analyze Peripheral Blood Mononuclear Cells in Chronic Chagas Disease

  • John E. Wiktorowicz
  • M. Paola Zago
  • Nisha J. GargEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1955)

Abstract

The thiol moieties of cysteinyl residues in proteins undergo a number of modifications including nitrosylation, oxidation, persulfidation, sulfenylation, and others. These protein modifications may influence gain as well as loss of function in biological and disease conditions. Herein, we describe a quantitative approach that combines accurate, sensitive fluorescence modification of cysteinyl-S-nitrosyl (SNOFlo) groups that leaves electrophoretic mobility unaffected and offers the measurement of changes in S-nitrosylation (SNO) status relative to protein abundance. This approach has been useful in evaluating the global protein abundance and SNO profile of Chagas seropositive individuals that were categorized in clinically asymptomatic (C/A) and clinically symptomatic (C/S) subgroups and compared to normal healthy (N/H) controls. Through analyzing the proteome datasets with different bioinformatics and statistics tools, potential pathologic mechanisms in disease progression are identified. We also propose a panel of protein biomarkers that have a potential to identify the infected individuals at risk of developing clinical Chagas disease.

Key words

Chagas cardiomyopathy Trypanosoma cruzi S-nitrosylation Peripheral blood mononuclear cells 2DE Mass spectrometry 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • John E. Wiktorowicz
    • 1
    • 4
  • M. Paola Zago
    • 2
  • Nisha J. Garg
    • 3
    • 4
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyUniversity of Texas Medical Branch (UTMB)GalvestonUSA
  2. 2.Instituto de Patología Experimental, Universidad Nacional de Salta – Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)SaltaArgentina
  3. 3.Department of Microbiology and ImmunologyUTMBGalvestonUSA
  4. 4.Institute for Human Infections and Immunity, UTMBGalvestonUSA

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