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Investigation of the plasmodium falciparum food vacuole through inducible expression of the chloroquine resistance transporter (PfCRT)
journal contribution
posted on 2012-06-13, 00:00 authored by F Ehlgen, J Pham, Tania De Koning-WardTania De Koning-Ward, A Cowman, S RalphHaemoglobin 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.
History
Journal
PLoS OneVolume
7Issue
6Publisher
Public Library of ScienceLocation
San Francisco, Calif.Publisher DOI
ISSN
1932-6203Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2012, Public Library of ScienceUsage metrics
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brefeldin Achloroquinechloroquine resistance transporterdynamingreen fluorescent proteinhemozoinmembrane proteinunclassified drugScience & TechnologyMultidisciplinary SciencesScience & Technology - Other TopicsTRANSMEMBRANE PROTEIN PFCRTDYNAMIN-LIKE PROTEINMALARIA PARASITEDIGESTIVE-VACUOLEENDOPLASMIC-RETICULUMGUANINE-NUCLEOTIDEHEMOGLOBIN UPTAKEDRUG-RESISTANCEMAURERS CLEFTSBREFELDIN-A
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