The human malaria parasite Plasmodium falciparum exports the ATP-binding cassette protein PFGCN20 to membrane structures in the host red blood cell.
PFGCN20 is a member of the ATP-binding cassette family of proteins that is closely related to the yeast translational regulator Gcn20p. We have generated a polyclonal antibody against the N-terminal region of PFGCN20 and studied the cellular localization of PFGCN20 throughout the erythrocytic life cycle of Plasmodium falciparum. PFGCN20 was found to be present at all stages and a pronounced export of PFGCN20 into the erythrocyte was observed in the trophozoite and schizont stages. In the indirect immunofluorescence assay, PFGCN20 was found to display significant colocalization with antigens detected by the monoclonal antibody 41E11. In contrast, there was only a minimal overlap of PFGCN20 localization with EMP2 and HRP2. Immunoelectron microscopy demonstrated a pronounced accumulation of PFGCN20 in the lumen of the parasitophorous vacuole and deconvolution fluorescence microscopy showed membrane association with selective regions of a tubovesicular network in the red cell. We also observed a concentration of PFGCN20 in electron-dense plaques just underneath the parasite's plasma membrane and an association of PFGCN20 with cytoplasmic vesicular structures within the parasite. The observed export of PFGCN20 and its association with the tubovesicular network in host red cells, may be indicative of the fact that PFGCN20 functions as ATP-binding subunit of an unknown multimeric ABC-transporter. The cytoplasmic localization of PFGCN20 in the parasite, however, suggests that the involvement of PFGCN20 in translational regulation or other cytoplasmic biological functions cannot be ruled out.
Bozdech
Z
Z
McGill Centre for the Study of Host Resistance, Department of Experimental Medicine, McGill University, Montreal, Quebec, Canada.
VanWye
J
J
Haldar
K
K
Schurr
E
E
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism* / Q000469:parasitology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting; D008018:Life Cycle Stages / Q000502:physiology; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D010963:Plasmodium falciparum / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D011506:Proteins / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism*; D000099053:Histidine-Rich Glycoprotein
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; C486403:EMP2 protein, human; C113388:Emp2 protein, mouse; D008562:Membrane Glycoproteins; C117857:PFGCN20 protein, Plasmodium falciparum; D011506:Proteins; D015800:Protozoan Proteins; D000099053:Histidine-Rich Glycoprotein