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Phosphoproteomic analysis of Leishmania donovani pro‐ and amastigote stages
Author(s) -
Morales Miguel A.,
Watanabe Reiko,
Laurent Christine,
Lenormand Pascal,
Rousselle JeanClaude,
Namane Abdelkader,
Späth Gerald F.
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700697
Subject(s) - amastigote , leishmania donovani , leishmania , biology , phosphoprotein , phosphorylation , microbiology and biotechnology , proteomics , biochemistry , kinase , phosphoproteomics , protein phosphorylation , intracellular parasite , intracellular , visceral leishmaniasis , parasite hosting , protein kinase a , leishmaniasis , gene , immunology , world wide web , computer science
Following transmission to the vertebrate host, the protozoan parasite Leishmania donovani differentiates into the pathogenic amastigote stage that is adapted for intracellular survival. This developmental transition is induced by environmental factors including elevated temperature and acidic pH and is likely transduced by signaling cascades involving protein kinases and their downstream phosphoprotein substrates. These signaling networks are highly adapted to the specific nutritional and physiological requirements of the organism and thus studying Leishmania phosphorylation may allow important insight into the parasite‐specific biology. We used a gel‐based approach to investigate qualitative and quantitative changes of the phosphoproteome of the major L. donovani life cycle stages. Phosphoproteins were purified by immobilized metal affinity chromatography (IMAC), separated by IEF and SDS‐PAGE using pH 4–7 IPG immobiline strips, revealed by fluorescent multiplex staining, and identified by MALDI‐MS and MS/MS. Our analysis allowed us to establish a first repertoire of the Leishmania phosphoproteome and to identify phosphoproteins implicated in stress‐ and heat shock response, RNA/protein turnover, metabolism, and signaling.