Phosphoproteomic analysis of Leishmania donovani pro- and amastigote stages.
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.
Morales
Miguel A
MA
Department of Parasitology and Mycology, Laboratory of Parasite Virulence, Institut Pasteur, 25-28 rue du Dr. Roux, Paris Cedex 15, France.
Watanabe
Reiko
R
Laurent
Christine
C
Lenormand
Pascal
P
Rousselle
Jean-Claude
JC
Namane
Abdelkader
A
Späth
Gerald F
GF
D000818:Animals; D002454:Cell Differentiation; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D007893:Leishmania donovani / Q000254:growth & development*; D013058:Mass Spectrometry; D010750:Phosphoproteins / Q000032:analysis*; D010766:Phosphorylation; D020033:Protein Isoforms / Q000032:analysis; D020543:Proteome; D015800:Protozoan Proteins / Q000032:analysis*
D010750:Phosphoproteins; D020033:Protein Isoforms; D020543:Proteome; D015800:Protozoan Proteins