Molecular characterization of a 2-Cys peroxiredoxin from the human malaria parasite Plasmodium falciparum

Kawazu, Shin Ichiro; Komaki, Kanako; Tsuji, Naotoshi; Kawai, Shinya; Ikenoue, Nozomu; Hatabu, Toshimitsu; Ishikawa, Hiroyuki; Matsumoto, Yoshitsugu; Himeno, Kunisuke; Kano, Shigeyuki

doi:10.1016/s0166-6851(01)00308-5

Cited by 52 publications

(39 citation statements)

References 30 publications

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“…For most of them, 1 to 16 M recombinant protein is necessary for complete DNA protection. It was shown that 5 M rLdmPrx inhibits nicking of DNA completely, which is in good correlation with the properties of the other peroxiredoxins (14,23,24,27,36).…”

Section: Resultsmentioning

confidence: 65%

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Expression of a Mitochondrial Peroxiredoxin Prevents Programmed Cell Death inLeishmania donovani

Harder

Bente

Isermann³

et al. 2006

Eukaryot Cell

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Leishmania promastigote cells transmitted by the insect vector get phagocytosed by macrophages and convert into the amastigote form. During development and transformation, the parasites are exposed to various concentrations of reactive oxygen species, which can induce programmed cell death (PCD). We show that a mitochondrial peroxiredoxin (LdmPrx) protects Leishmania donovani from PCD. Whereas this peroxiredoxin is restricted to the kinetoplast area in promastigotes, it covers the entire mitochondrion in amastigotes, accompanied by dramatically increased expression. A similar change in the expression pattern was observed during the growth of Leishmania from the early to the late logarithmic phase. Recombinant LdmPrx shows typical peroxiredoxin-like enzyme activity. It is able to detoxify organic and inorganic peroxides and prevents DNA from hydroxyl radical-induced damage. Most notably, Leishmania parasites overexpressing this peroxiredoxin are protected from hydrogen peroxide-induced PCD. This protection is also seen in promastigotes grown to the late logarithmic phase, also characterized by high expression of this peroxiredoxin. Apparently, the physiological role of this peroxiredoxin is stabilization of the mitochondrial membrane potential and, as a consequence, inhibition of PCD through removal of peroxides.Leishmania parasites affect more than 12 million people worldwide, with an estimated 2 million new cases each year (WHO World Health Report, 2004, http://www.who.int/whr /en). Depending on the species involved, symptoms range from the self-healing cutaneous form (Leishmania major) to the fatal visceral form (L. donovani). The parasite is transmitted as the infective promastigote form from the gut of its insect vector, female phlebotomine flies of the genera Phlebotomus and Lutzomyia, to mammalian hosts. Promastigotes get phagocytosed by macrophages and convert into the amastigote form, which is able to survive and replicate within phagolysosomes. During phagocytosis of Leishmania promastigotes, the macrophages produce different reactive oxygen species (ROS) to kill the parasites. ROS readily react with proteins, DNA, and lipids and have been implicated in a wide variety of cell functions, such as signal transduction, redox homeostasis, apoptosis, aging, tumor progression, and pathogen infection (9,19,42,58). Numerous reports have shown that Leishmania parasites are susceptible to ROS-and RNS (reactive nitrogen species)-mediated toxicity (41, 57). In order to survive and establish an infection, they have to cope with these pro-oxidants. In Trypanosomatidae, it was shown that peroxiredoxins are the major antioxidant enzymes that can use different ROS and RNS like H 2 O 2 , hydroperoxides, and ONOO as substrates (56). Peroxiredoxins are found in a great variety of organisms, where they fulfill distinct functions, such as detoxification, signaling, or differentiation (25). In different members of the family Trypanosomatidae, cytosolic, as well as mitochondrial, peroxiredoxins were found (6,10,11,21). Peroxire...

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Section: Resultsmentioning

confidence: 65%

“…The enzymatic kinetics observed for the removal of H 2 O 2 is comparable to that of peroxiredoxins from P. falciparum, C. elegans, human neutrophils, and Pisum sativum, where similar peroxidase assays were used (Fig. 1D) (8,26,27,32).…”

Section: Resultsmentioning

confidence: 67%

Expression of a Mitochondrial Peroxiredoxin Prevents Programmed Cell Death inLeishmania donovani

Harder

Bente

Isermann³

et al. 2006

Eukaryot Cell

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“…Prx protein from Plasmodium constitutes about 0.25% of the total protein in late-stage trophozoites and is constitutively expressed during asexual development as well as in the gametocyte, both in P. falciparum [16] and rodent Plasmodium parasites [17,18]. Since this protein is present in the cytoplasm of malarial parasites but is not associated with the parasite membrane, it is likely that it might be released from the parasites when schizonts are ruptured to release merozoites, or when erythrocytes infected with malarial parasites are destroyed in the spleen.…”

Section: Discussionmentioning

confidence: 99%

“…Recombinant 2-Cys PbAPrx or PfPrx was produced in E. coli and purified as described previously [16,17]. The purity of the recombinant protein was confirmed by SDS-PAGE (data not shown).…”

Section: Production Of Recombinant Malarial Prxmentioning

confidence: 99%

Mast cell‐mediated immune responses through IgE antibody and Toll‐like receptor 4 by malarial peroxiredoxin

et al. 2008

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In this study, 2-Cys Plasmodium berghei ANKA (PbA) peroxiredoxin (Prx) was identified as an antigenic protein recognized by an anti-PbA IgE antibody using two-dimensional polyacrylamide gel electrophoresis and proteomic analysis. Innate immune responses to PbAPrx were examined using cells from mice deficient in Toll-like receptors (TLR) or related molecules, and it was demonstrated that responses were severely impaired in TLR4 -/-, MyD88 -/-and MD-2 -/-mice, but not in Toll/IL-1 receptor domain-containing adaptor inducing IFN-c (TRIF) -/-, TLR2 -/-or radioprotective 105 (RP105) -/-mice. An association between PbAPrx and TLR4 was observed following immunoprecipitation and immunoblotting, suggesting that PbAPrx was associated with TLR4/MD-2. Interactions between Prx and TLR4/MD-2 were also examined by flow cytometry using TLR4/MD-2-or TLR2-expressing cells. NFjB/GFP activity was observed in TLR4/MD-2-but not in TLR2-expressing cells following stimulation with Prx. However, this effect was not observed after treatment with proteinase K, suggesting that PbAPrx is a protein ligand for TLR4 and that the PbAPrx activity observed in this study is not due to contamination with LPS. These findings indicate that malarial Prx induces IgE-mediated protection through FceRI on mast cells and innate immunity through TLR4 with MyD88 and MD-2, suggesting a novel function for malarial Prx in innate and acquired immune responses in malaria.

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“…Trx is reduced by TrxR, a flavoenzyme with high structural and mechanistic homology to GR. A glutathione-dependent peroxidase seems to be absent in P. falciparum, whereas at least four Trx-dependent peroxidases appear to be present (19,23,24). Both the glutathione system, as indicated by drug studies (25), and the Trx system, as shown by TrxR knock-outs (26), are essential for parasite viability.…”

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confidence: 99%

Identification and Characterization of Heme-interacting Proteins in the Malaria Parasite, Plasmodium falciparum

Campanale

Nickel²,

Daubenberger

et al. 2003

Journal of Biological Chemistry

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The degradation of hemoglobin by the malaria parasite, Plasmodium falciparum, produces free ferriprotoporphyrin IX (FP) as a toxic by-product. In the presence of FP-binding drugs such as chloroquine, FP detoxification is inhibited, and the build-up of free FP is thought to be a key mechanism in parasite killing. In an effort to identify parasite proteins that might interact preferentially with FP, we have used a mass spectrometry approach. Proteins that bind to FP immobilized on agarose include P. falciparum glyceraldehyde-3-phosphate dehydrogenase (PfGAPDH), P. falciparum glutathione reductase (PfGR), and P. falciparum protein disulfide isomerase. To examine the potential consequences of FP binding, we have examined the ability of FP to inhibit the activities of GAPDH and GR from P. falciparum and other sources. FP inhibits the enzymic activity of Pf-GAPDH with a K i value of 0.2 M, whereas red blood cell GAPDH is much less sensitive. By contrast, PfGR is more resistant to FP inhibition (K i > 25 M) than its human counterpart. We also examined the ability of FP to inhibit the activities of the additional antioxidant enzymes, P. falciparum thioredoxin reductase, which exhibits a K i value of 1 M, and P. falciparum glutaredoxin, which shows more moderate sensitivity to FP. The exquisite sensitivity of PfGAPDH to FP may indicate that the glycolytic pathway of the parasite is particularly susceptible to modulation by FP stress. Inhibition of this pathway may drive flux through the pentose phosphate pathway ensuring sufficient production of reducing equivalents to counteract the oxidative stress induced by FP build-up.

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Molecular characterization of a 2-Cys peroxiredoxin from the human malaria parasite Plasmodium falciparum

Cited by 52 publications

References 30 publications

Expression of a Mitochondrial Peroxiredoxin Prevents Programmed Cell Death inLeishmania donovani

Expression of a Mitochondrial Peroxiredoxin Prevents Programmed Cell Death inLeishmania donovani

Mast cell‐mediated immune responses through IgE antibody and Toll‐like receptor 4 by malarial peroxiredoxin

Identification and Characterization of Heme-interacting Proteins in the Malaria Parasite, Plasmodium falciparum

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