Plasmodium falciparum proteome changes in response to doxycycline treatment.
BACKGROUND
The 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.


METHODS
A 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).


RESULTS
After 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.


CONCLUSIONS
In 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.
Briolant
Sébastien
S
Unité de Recherche en Biologie et Epidémiologie Parasitaires, Institut de Recherche Biomédicale des Armées, Antenne de Marseille, BP 60109, 13262 Marseille Cedex 07, France.
Almeras
Lionel
L
Belghazi
Maya
M
Boucomont-Chapeaublanc
Elodie
E
Wurtz
Nathalie
N
Fontaine
Albin
A
Granjeaud
Samuel
S
Fusaï
Thierry
T
Rogier
Christophe
C
Pradines
Bruno
B
D000962:Antimalarials / Q000494:pharmacology*; D004318:Doxycycline / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation / Q000187:drug effects; D017125:Genes, Protozoan; D023281:Genomics; D006801:Humans; D016778:Malaria, Falciparum / Q000188:drug therapy / Q000469:parasitology; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D010641:Phenotype; D010963:Plasmodium falciparum / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D015800:Protozoan Proteins / Q000187:drug effects / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D052939:Schizonts / Q000737:chemistry / Q000378:metabolism*
D000962:Antimalarials; D020543:Proteome; D015800:Protozoan Proteins; D004318:Doxycycline