Journal
MOLECULAR AND BIOCHEMICAL PARASITOLOGY
Volume 158, Issue 2, Pages 189-201Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.molbiopara.2007.12.008
Keywords
Trypanosoma brucei; proteomics; glycosome; mitochondrion; glycolysis; metabolism; cell fractionation; liquid chromatography; mass spectrometry
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Label-free semi-quantitative differential three-dimensional liquid chromatography coupled to mass spectrometry (3D-LC-MS/MS) was used to compare the glycosomal and mitochondrial proteomes of the bloodstream- and insect-form of Trypanosoma brucei. The abundance of glycosomal marker proteins identified in the two life-cycle stages corresponded well with the relative importance of biochemical pathways present in the glycosomes of the two stages and the peptide spectral count ratios of selected enzymes were in good agreement with published data about their enzymatic specific activities. This approach proved extremely useful for the generation of large scale proteomics data for the comparison of different life-cycle stages. Several proteins involved in oxidative stress protection, sugar-nucleotide synthesis, purine salvage, nucleotide-monophosphate formation and purine-nucleotide cycle were identified as glycosomal proteins. (c) 2007 Elsevier B.V. All rights reserved.
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