Proteome remodelling during development from blood to insect-form Trypanosoma brucei quantified by SILAC and mass spectrometry.
BACKGROUND
Trypanosoma brucei is the causative agent of human African sleeping sickness and Nagana in cattle. In addition to being an important pathogen T. brucei has developed into a model system in cell biology.


RESULTS
Using Stable Isotope Labelling of Amino acids in Cell culture (SILAC) in combination with mass spectrometry we determined the abundance of >1600 proteins in the long slender (LS), short stumpy (SS) mammalian bloodstream form stages relative to the procyclic (PC) insect-form stage. In total we identified 2645 proteins, corresponding to ~30% of the total proteome and for the first time present a comprehensive overview of relative protein levels in three life stages of the parasite.


CONCLUSIONS
We can show the extent of pre-adaptation in the SS cells, especially at the level of the mitochondrial proteome. The comparison to a previously published report on monomorphic in vitro grown bloodstream and procyclic T. brucei indicates a loss of stringent regulation particularly of mitochondrial proteins in these cells when compared to the pleomorphic in vivo situation. In order to better understand the different levels of gene expression regulation in this organism we compared mRNA steady state abundance with the relative protein abundance-changes and detected moderate but significant correlation indicating that trypanosomes possess a significant repertoire of translational and posttranslational mechanisms to regulate protein abundance.
Institute for Cell Biology, University of Bern, Bern, Switzerland.
Kapila
K
Gunasekera
Daniel
D
Wüthrich
Sophie
S
Braga-Lagache
Manfred
M
Heller
Torsten
T
Ochsenreiter
D017398:Alternative Splicing / Q000235:genetics; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019295:Computational Biology; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D007553:Isotope Labeling; D008018:Life Cycle Stages / Q000235:genetics*; D013058:Mass Spectrometry; D024101:Mitochondrial Proteins / Q000378:metabolism*; D025341:Principal Component Analysis; D020543:Proteome / Q000235:genetics*; D014346:Trypanosoma brucei brucei / Q000235:genetics / Q000254:growth & development*
D000596:Amino Acids; D024101:Mitochondrial Proteins; D020543:Proteome