Investigation of the Plasmodium falciparum Food Vacuole through Inducible Expression of the Chloroquine Resistance Transporter (PfCRT)
- Author(s)
- Ehlgen, F; Pham, JS; de Koning-Ward, T; Cowman, AF; Ralph, SA;
- Details
- Publication Year 2012-06-13,Volume 7,Issue #6,Page e38781
- Journal Title
- PLOS ONE
- Publication Type
- Journal Article
- Abstract
- 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.
- Publisher
- PUBLIC LIBRARY SCIENCE
- Keywords
- TRANSMEMBRANE PROTEIN PFCRT; DYNAMIN-LIKE PROTEIN; MALARIA PARASITES; DIGESTIVE-VACUOLE; ENDOPLASMIC-RETICULUM; INFECTED ERYTHROCYTES; GUANINE-NUCLEOTIDE; HEMOGLOBIN UPTAKE; DRUG RESISTANCE; MAURERS CLEFTS
- Research Division(s)
- Infection And Immunity
- Publisher's Version
- https://doi.org/10.1371/journal.pone.0038781
- Open Access at Publisher's Site
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3374814/- Terms of Use/Rights Notice
- Copyright: © 2012 Ehlgen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Creation Date: 2012-06-13 12:00:00