Document Detail

Investigation of the Plasmodium falciparum food vacuole through inducible expression of the chloroquine resistance transporter (PfCRT).
MedLine Citation:
PMID:  22719945     Owner:  NLM     Status:  MEDLINE    
Haemoglobin degradation during the erythrocytic life stages is the major function of the food vacuole (FV) of Plasmodium falciparum and the target of several anti-malarial drugs that interfere with this metabolic pathway, killing the parasite. Two multi-spanning food vacuole membrane proteins are known, the multidrug resistance protein 1 (PfMDR1) and Chloroquine Resistance Transporter (PfCRT). Both modulate resistance to drugs that act in the food vacuole. To investigate the formation and behaviour of the food vacuole membrane we have generated inducible GFP fusions of chloroquine sensitive and resistant forms of the PfCRT protein. The inducible expression system allowed us to follow newly-induced fusion proteins, and corroborated a previous report of a direct trafficking route from the ER/Golgi to the food vacuole membrane. These parasites also allowed the definition of a food vacuole compartment in ring stage parasites well before haemozoin crystals were apparent, as well as the elucidation of secondary PfCRT-labelled compartments adjacent to the food vacuole in late stage parasites. We demonstrated that in addition to previously demonstrated Brefeldin A sensitivity, the trafficking of PfCRT is disrupted by Dynasore, a non competitive inhibitor of dynamin-mediated vesicle formation. Chloroquine sensitivity was not altered in parasites over-expressing chloroquine resistant or sensitive forms of the PfCRT fused to GFP, suggesting that the PfCRT does not mediate chloroquine transport as a GFP fusion protein.
Florian Ehlgen; James S Pham; Tania de Koning-Ward; Alan F Cowman; Stuart A Ralph
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-06-13
Journal Detail:
Title:  PloS one     Volume:  7     ISSN:  1932-6203     ISO Abbreviation:  PLoS ONE     Publication Date:  2012  
Date Detail:
Created Date:  2012-06-21     Completed Date:  2012-12-13     Revised Date:  2013-07-12    
Medline Journal Info:
Nlm Unique ID:  101285081     Medline TA:  PLoS One     Country:  United States    
Other Details:
Languages:  eng     Pagination:  e38781     Citation Subset:  IM    
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, Victoria, Australia.
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MeSH Terms
Base Sequence
Blotting, Western
Brefeldin A / pharmacology
Cell Compartmentation
Chloroquine / pharmacology
DNA Primers
Fluorescent Antibody Technique
Membrane Transport Proteins / metabolism*
Plasmodium falciparum / metabolism*
Polymerase Chain Reaction
Protozoan Proteins / metabolism*
Vacuoles / metabolism*
Reg. No./Substance:
0/DNA Primers; 0/Membrane Transport Proteins; 0/PfCRT protein, Plasmodium falciparum; 0/Protozoan Proteins; 20350-15-6/Brefeldin A; 54-05-7/Chloroquine

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