Large-scale differential proteome analysis in Plasmodium falciparum under drug treatment.

PloS One
Judith Helena PrietoKatja Becker

Abstract

Proteome studies contribute markedly to our understanding of parasite biology, host-parasite interactions, and mechanisms of drug action. For most antimalarial drugs neither mode of action nor mechanisms of resistance development are fully elucidated although this would be important prerequisites for successfully developing urgently required novel antimalarials. Here, we establish a large-scale quantitative proteomic approach to examine protein expression changes in trophozoite stages of the malarial parasite Plasmodium falciparum following chloroquine and artemisinin treatment. For this purpose SIL (stable isotope labeling) using (14)N-isoleucine and (13)C(6),(15)N(1)-isoleucine was optimized to obtain 99% atomic percent 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 novel software tool Census. In drug treated parasites, we identified a total number of 1,253 proteins, thus increasing the overall number of proteins identified in the trophozoite stage by 30%. A relative quantification was obtained f...Continue Reading

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Citations

Jun 29, 2011·Future Microbiology·Case McNamara, Elizabeth A Winzeler
Aug 12, 2009·Proceedings of the National Academy of Sciences of the United States of America·Sasa KoncarevicKatja Becker
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Methods Mentioned

BETA
electrophoresis
ion exchanger

Software Mentioned

Census
PlasmoDB
DTASelect2
MudPIT
Quantity One
SEQUEST
DTASelect

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