Post-translational modification of cellular proteins during Leishmania donovani differentiation.
The pathogenic intracellular parasites Leishmania donovani cycle between sand fly gut and the human macrophage phagolysosome, differentiating from extracellular promastigotes to intracellular amastigote forms. Using isobaric tagging for relative and absolute quantifications (iTRAQ/LC-MS/MS) proteomic methodology, we recently described the ordered gene expression changes during this process. While protein abundance changes in Leishmania were documented, little is known about their PTMs. Here we used iTRAQ to detect protein phosphorylation, methylation, acetylation, and glycosylation sites throughout differentiation. We found methylation of arginines, aspartic acids, glutamic acids, asparagines, and histidines. Detected acetylation sites included serines and protein N-terminal acetylations on methionines, serines, alanines, and threonines. Phosphorylations were detected on serines and threonines, but not tyrosines. iTRAQ identified novel fucosylation sites as well as hexosylations. We observed quantity changes in some modifications during differentiation, suggesting a role in L. donovani intracellular development. This study is the first high-throughput analysis of PTM sites dynamics during an intracellular parasitic development.
Rosenzweig
Doron
D
Faculty of Biology, Technion - Israel Institute of Technology, Haifa, Israel.
Smith
Derek
D
Myler
Peter J
PJ
Olafson
Robert W
RW
Zilberstein
Dan
D
D000818:Animals; D001216:Asparagine / Q000737:chemistry; D001224:Aspartic Acid / Q000737:chemistry; D002454:Cell Differentiation; D002853:Chromatography, Liquid / Q000379:methods; D018698:Glutamic Acid / Q000737:chemistry; D006031:Glycosylation; D006639:Histidine / Q000737:chemistry; D006801:Humans; D007893:Leishmania donovani / Q000378:metabolism*; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology*; D013058:Mass Spectrometry / Q000379:methods; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*
D001224:Aspartic Acid; D018698:Glutamic Acid; D006639:Histidine; D001216:Asparagine