Whole Proteome Analysis of the Protozoan Parasite Trypanosoma brucei using Stable Isotope Labeling by Amino Acids in Cell Culture and Mass Spectrometry

Cirovic, Olivera; Ochsenreiter, Torsten (2013). Whole Proteome Analysis of the Protozoan Parasite Trypanosoma brucei using Stable Isotope Labeling by Amino Acids in Cell Culture and Mass Spectrometry. In: SILAC. Methods in Molecular Biology. Heidelberg: Springer

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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 allowed for the characterization of the proteome from several different life stages of the parasite (1-3). To achieve this, stable isotope labeling by amino acids in cell culture (SILAC; (4)) 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 knock-out approaches.

Item Type:

Book Section (Book Chapter)

Division/Institute:

08 Faculty of Science > Other Institutions > Teaching Staff, Faculty of Science
08 Faculty of Science > Department of Biology > Institute of Cell Biology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Cirovic, Olivera and Ochsenreiter, Torsten

Subjects:

500 Science > 570 Life sciences; biology

Series:

Methods in Molecular Biology

Publisher:

Springer

Funders:

[4] Swiss National Science Foundation

Language:

English

Submitter:

Torsten Ochsenreiter

Date Deposited:

15 Apr 2014 12:01

Last Modified:

10 Aug 2015 16:10

URI:

https://boris.unibe.ch/id/eprint/45778

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