Premium
Analysis of Trypanosoma cruzi Phosphoproteome by Mass Spectrometry
Author(s) -
Gaynor Matthew R,
Nakayasu Ernesto S,
Almeida Igor C
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1026.4
Subject(s) - trypanosoma cruzi , chagas disease , phosphorylation , genbank , proteomics , signal transduction , biology , chemistry , biochemistry , protein phosphorylation , computational biology , microbiology and biotechnology , protein kinase a , parasite hosting , gene , virology , world wide web , computer science
Trypanosoma cruzi is the etiologic agent of Chagas’ disease, which affects over 11 million people in Latin America. Lately, much research has been focused on drug and vaccine development against this parasite. In eukaryotic cells, protein phosphorylation regulates many vital pathways and has been extensively studied as potential molecular target. Here, we analyzed T. cruzi phosphoproteins and mapped the phosphorylation sites through mass spectrometry (MS). Total proteins from T. cruzi epimastigote forms were digested with trypsin/urea and fractionated in a strong cation‐exchange ziptip. Ensuing phosphopeptides were purified by immobilized metal‐affinity chromatography (IMAC) and analyzed by liquid chromatography‐tandem MS (LC‐MS/MS, ‐MS3, ‐pseudoMS3). The resulting spectra were searched against T. cruzi sequences in the GenBank using Sequest algorithm. We were able to identify 143 phosphorylation sites from 98 distinct proteins. The Gene Ontology analysis revealed several proteins with protein‐ and nucleotide‐binding activity, cytoskeleton proteins, and proteins involved signal transduction. To our best knowledge, this is the first phosphoproteome study performed in a pathogenic protozoan. Our data may provide new insights into discovery of new interventions to prevent or treat Chagas’ disease. Supported by NSF/REU Program (grant#DBI‐0353887), and NIH/SCORE (grant#2SO6GM0812‐37).