Plasmodium falciparum proteome changes in response to doxycycline treatment

Abstract

BackgroundThe emergence of Plasmodium falciparum resistance to most anti-malarial compounds has highlighted the urgency to develop new drugs and to clarify the mechanisms of anti-malarial drugs currently used. Among them, doxycycline is used alone for malaria chemoprophylaxis or in combination with quinine or artemisinin derivatives for malaria treatment. The molecular mechanisms of doxycycline action in P. falciparum have not yet been clearly defined, particularly at the protein level.MethodsA proteomic approach was used to analyse protein expression changes in the schizont stage of the malarial parasite P. falciparum following doxycycline treatment. A comparison of protein expression between treated and untreated protein samples was performed using two complementary proteomic approaches: two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and isobaric tagging reagents for relative and absolute quantification (iTRAQ).ResultsAfter doxycycline treatment, 32 and 40 P. falciparum proteins were found to have significantly deregulated expression levels by 2D-DIGE and iTRAQ methods, respectively. Although some of these proteins have been already described as being deregulated by other drug treatments, numerous changes in protein levels seem to be specific to doxycycline treatment, which could perturb apicoplast metabolism. Quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to confirm this hypothesis.ConclusionsIn this study, a specific response to doxycycline treatment was distinguished and seems to involve mitochondrion and apicoplast organelles. These data provide a starting point for the elucidation of drug targets and the discovery of mechanisms of resistance to anti-malarial compounds.

Quantitative Time-course Profiling of Parasite and Host Cell Proteins in the Human Malaria Parasite Plasmodium falciparum

“…has been shown to be up-regulated on a translational but not on a transcriptional level following drug treatment (10). Other studies have evaluated the links between transcription and translation during the Plasmodium life cycle progression in more general terms by employing different types of large scale proteomics assays (7,(11)(12)(13)(14)(15)(16)(17)(18)(19). These studies thus complemented the transcriptome analyses and demonstrated that each developmental stage of Plasmodium is characterized also by a unique protein content that is linked to the specialized functionalities of that particular stage.…”

Caracterización parcial del proteoma del trofozoito de Plasmodium falciparum bajo tratamiento con quinina, mefloquina y el antiplasmodial natural diosgenona

“…The apparent differential expression of 10 kDa chaperonin in response to mefloquine and its localization deserve further investigation. The differential expression of the chaperonin proteins under different drug treatments in P. falciparum has been reported previously (4,10).…”

“…Enolase has been identified as an abundant protein in twodimensional gels (6). The differential expression of enolase has been reported in the parasite under pressure with artemether-lumefatrine (6) and doxycycline (10). However, this was not observed when the parasite was treated with pyrimethamine (7).…”

“…Protein spots with p < 0.05 were considered differentially expressed. Image Scanner III (GE Healthcare) was previously calibrated as described (10).…”

“…Proteomic approaches have allowed the understanding of the molecular events occurring in the parasite in response to different antimalarial drugs and environments. However, some are based on different proteomic techniques with contrasting results (4)(5)(6)(7)(8)(9)(10)(11).…”