title: Histidine-rich protein 2 of the malaria parasite, Plasmodium falciparum, is involved in detoxification of the by-products of haemoglobin degradation.
abstract: The histidine-rich protein 2 (PfHRP2) of Plasmodium falciparum has been implicated in the detoxification of ferriprotoporphyrin IX (FP) moieties that are produced as by-products of the digestion of haemoglobin. In this work, we have used a spectroscopic analysis to confirm that recombinant PfHRP2 binds FP. A monoclonal antibody that recognises both recombinant and authentic PfHRP2 was used in immunofluorescence microscopy studies. We found that PfHRP2 is mainly located in the erythrocyte cytosol of infected erythrocytes, however, dual labelling studies suggest that the location of a sub-population of the PfHRP2 molecules overlaps with that of the food vacuole-associated protein, P-glycoprotein homologue (Pgh-1). A semi-quantitative analysis of the level of PfHRP2 in infected erythrocytes suggests a concentration of a few micromolar in the food vacuole. Under conditions designed to mimic the parasite food vacuole, we found that 1.2 microM PfHRP2 is sufficient to catalyse the conversion of about 30% of a 100 microM sample of FP to beta-haematin within 24 h. Moreover, PfHRP2 is capable of promoting the H(2)O(2)-induced degradation of FP at pH 5.2. PfHRP2 also efficiently enhances the ability of FP to catalyse the H(2)O(2)-mediated oxidation of the model co-factor, ortho-phenylene diamine (OPD). These data suggest that PfHRP2 may promote the detoxification of FP and reactive oxygen species within the food vacuole. By contrast, PfHRP2 inhibits the destruction of FP by glutathione (GSH) at pH 7.4. This suggests that PfHRP2 is not a catalyst of FP degradation outside the food vacuole.
authors.affiliation: Department of Biochemistry, La Trobe University, Bundoora, 3083, Victoria, Australia.
authors.forename: V
authors.initials: V
authors.lastname: Papalexis
authors.forename: M A
authors.initials: MA
authors.lastname: Siomos
authors.forename: N
authors.initials: N
authors.lastname: Campanale
authors.forename: X
authors.initials: X
authors.lastname: Guo
authors.forename: G
authors.initials: G
authors.lastname: Kocak
authors.forename: M
authors.initials: M
authors.lastname: Foley
authors.forename: L
authors.initials: L
authors.lastname: Tilley
mesh_terms: D000818:Animals; D004912:Erythrocytes / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D006427:Hemin / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism; D006801:Humans; D016778:Malaria, Falciparum; D051379:Mice; D018613:Microscopy, Confocal; D010084:Oxidation-Reduction; D010963:Plasmodium falciparum / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D000099053:Histidine-Rich Glycoprotein
chemical_list: D006454:Hemoglobins; D011506:Proteins; D000099053:Histidine-Rich Glycoprotein; D006427:Hemin