Sammelbandbeitrag

Erschienen in: Parasite Genomics Protocols

Herausgeberliste: Peacock, C

Jahr der Veröffentlichung: 2015

Seiten: 269-279

ISBN: 978-1-4939-1437-1

eISBN: 978-1-4939-1438-8

DOI Link: https://doi.org/10.1007/978-1-4939-1438-8_17

Auflage: 2. edition

Serientitel: Methods in molecular biology

Serienzählung: 1201

Abstract: 

Here, we establish a methodology for large-scale quantitative proteomics using SIL (stable isotope labeling) to examine protein expression changes in trophozoite stages of the malaria parasite Plasmodium falciparum following drug treatment. For this purpose, exposure to ¹³C₆ ¹⁵N₁-isoleucine was optimized in order to obtain 99 % atomic enrichment. Proteome fractionation with anion exchange chromatography was used to reduce sample complexity and increase quantitative coverage of protein expression. Tryptic peptides of subfractions were subjected to SCX/RP separation, measured by LC-MS/MS, and quantified using the software tool Census. In drug-treated parasites, we identified a total number of 1,253 proteins, thus increasing the overall number of proteins so far identified in the trophozoite stage by 30 % in the previous literature. A relative quantification was obtained for more than 800 proteins. About 5 % of proteins showed a clear up- or downregulation upon drug treatment.

Harvard-Zitierstil: Prieto, J., Fischer, E., Koncarevic, S., Yates, J. and Becker, K. (2015) Large-Scale Differential Proteome Analysis in Plasmodium falciparum Under Drug Treatment, in Peacock, C. (ed.) Parasite Genomics Protocols. 2. edition. New York: Humana Press, pp. 269-279. https://doi.org/10.1007/978-1-4939-1438-8_17

APA-Zitierstil: Prieto, J., Fischer, E., Koncarevic, S., Yates, J., & Becker, K. (2015). Large-Scale Differential Proteome Analysis in Plasmodium falciparum Under Drug Treatment. In Peacock, C. (Ed.), Parasite Genomics Protocols (2. edition, pp. 269-279). Humana Press. https://doi.org/10.1007/978-1-4939-1438-8_17