title: Whole proteome analysis of the protozoan parasite Trypanosoma brucei using stable isotope labeling by amino acids in cell culture and mass spectrometry.
abstract: The single-celled protozoan Trypanosoma brucei spp. is the causative agent of human African trypanosomiasis and nagana in cattle. Quantitative proteomics for the first time has allowed for the characterization of the proteome from several different life stages of the parasite (Butter et al., Mol Cell Proteomics 12:172-179, 2013; Gunasekera et al., BMC Genomics 13:556, 2012; Urbaniak et al., PloS One 7(5):e36619, 2012). To achieve this, stable isotope labeling by amino acids in cell culture (SILAC) (Ong et al., Mol Cell Proteomics 1:376-386, 2002) was adapted to T. brucei spp. cultures. T. brucei cells grown in standard media with dialyzed fetal calf serum containing heavy isotope-labeled amino acids (arginine and lysine) show efficient incorporation of the labeled amino acids into the whole cell proteome (8-12 divisions) and no detectable amino acid conversions. The method can be applied to both of the major life stages of the parasite and in combination with RNAi or gene knockout approaches.
authors: Institute of Cell Biology, University of Bern, Office C365, Baltzerstrasse 4, 3012, Bern, Switzerland.
authors: Olivera
authors: O
authors: Cirovic
authors: Torsten
authors: T
authors: Ochsenreiter
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D003956:Dialysis; D006358:Hot Temperature; D007553:Isotope Labeling / Q000379:methods*; D008018:Life Cycle Stages; D013058:Mass Spectrometry / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003470:Culture Media; D015800:Protozoan Proteins