Molecular cloning, characterization and expression profile of a glutathione peroxidase-like thioredoxin peroxidase (TPxGl) of the rodent malaria parasite Plasmodium berghei

Haselton, Kyle J.; David, Robin; Fell, Katherine; Schulte, Emily; Dybas, Matthew; Olsen, Kenneth W.; Kanzok, Stefan M.

doi:10.1016/j.parint.2014.02.004

Cited by 8 publications

(6 citation statements)

References 36 publications

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“…Plasmodium falciparum Tpx-1 knockout lines show an increased sensitivity to both ROS and reactive nitrogen species, but the deletion mutation is not lethal, suggesting redundancy in the parasite’s antioxidant system [22]. A glutathione peroxidase-like thioredoxin peroxidase (TPx Gl ) has also been shown to be part of the thioredoxin system in Plasmodium and is reduced by Trx-1 [23]. Inhibitors of the thioredoxin pathway have been investigated as potential anti-malarial compounds, rendering the parasite more susceptible to oxidative stress [24].…”

Section: Introductionmentioning

confidence: 99%

Functional genomic analyses of Enterobacter, Anopheles and Plasmodium reciprocal interactions that impact vector competence

Dennison

Saraiva

Cirimotich

et al. 2016

Malar J

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BackgroundMalaria exerts a tremendous socioeconomic impact worldwide despite current control efforts, and novel disease transmission-blocking strategies are urgently needed. The Enterobacter bacterium Esp_Z, which is naturally harboured in the mosquito midgut, can inhibit the development of Plasmodium parasites prior to their invasion of the midgut epithelium through a mechanism that involves oxidative stress. Here, a multifaceted approach is used to study the tripartite interactions between the mosquito, Esp_Z and Plasmodium, towards addressing the feasibility of using sugar-baited exposure of mosquitoes to the Esp_Z bacterium for interruption of malaria transmission.MethodsThe ability of Esp_Z to colonize Anopheles gambiae midguts harbouring microbiota derived from wild mosquitoes was determined by qPCR. Upon introduction of Esp_Z via nectar feeding, the permissiveness of colonized mosquitoes to Plasmodium falciparum infection was determined, as well as the impact of Esp_Z on mosquito fitness parameters, such as longevity, number of eggs laid and number of larvae hatched. The genome of Esp_Z was sequenced, and transcriptome analyses were performed to identify bacterial genes that are important for colonization of the mosquito midgut, as well as for ROS-production. A gene expression analysis of members of the oxidative defence pathway of Plasmodium berghei was also conducted to assess the parasite’s oxidative defence response to Esp_Z exposure.ResultsEsp_Z persisted for up to 4 days in the An. gambiae midgut after introduction via nectar feeding, and was able to significantly inhibit Plasmodium sporogonic development. Introduction of this bacterium did not adversely affect mosquito fitness. Candidate genes involved in the selection of a better fit Esp_Z to the mosquito midgut environment and in its ability to condition oxidative status of its surroundings were identified, and parasite expression data indicated that Esp_Z is able to induce a partial and temporary shutdown of the ookinetes antioxidant response.ConclusionsEsp_Z is capable of inhibiting sporogonic development of Plasmodium in the presence of the mosquito’s native microbiota without affecting mosquito fitness. Several candidate bacterial genes are likely mediating midgut colonization and ROS production, and inhibition of Plasmodium development appears to involve a shutdown of the parasite’s oxidative defence system. A better understanding of the complex reciprocal tripartite interactions can facilitate the development and optimization of an Esp_Z-based malaria control strategy.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1468-2) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Functional genomic analyses of Enterobacter, Anopheles and Plasmodium reciprocal interactions that impact vector competence

Dennison

Saraiva

Cirimotich

et al. 2016

Malar J

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“…PbPhLP-3 and HsPhLP-3 are reduced by the Trx-system in vitro and exhibit enzyme kinetics similar to those of other recently characterized Trx-like proteins in Plasmodium [7, 38, 47]. This interaction raises the possibility of PhLP-3 being a Trx-target protein in vivo .…”

Section: Discussionmentioning

confidence: 54%

“…berghei Trx-system, which consists of the P . berghei thioredoxin reductase (PbTrxR) and PbTrx-1 as previously described [38]. PbPhLP-3 was added to a reaction containing NADPH, and the P .…”

Section: Resultsmentioning

confidence: 99%

Characterization of a protozoan Phosducin-like protein-3 (PhLP-3) reveals conserved redox activity

et al. 2018

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We recently identified three novel thioredoxin-like genes in the genome of the protozoan parasite Plasmodium that belong to the Phosducin-like family of proteins (PhLP). PhLPs are small cytosolic proteins hypothesized to function in G-protein signaling and protein folding. Although PhLPs are highly conserved in eukaryotes from yeast to mammals, only a few representatives have been experimentally characterized to date. In addition, while PhLPs contain a thioredoxin domain, they lack a CXXC motif, a strong indicator for redox activity, and it is unclear whether members of the PhLP family are enzymatically active. Here, we describe PbPhLP-3 as the first phosducin-like protein of a protozoan organism, Plasmodium berghei. Initial transcription analysis revealed continuous low-level expression of pbphlp-3 throughout the complex Plasmodium life cycle. Attempts to knockout pbphlp-3 in P. berghei did not yield live parasites, suggesting an essential role for the gene in Plasmodium. We cloned, expressed and purified PbPhLP-3 and determined that the recombinant protein is redox active in vitro in a thioredoxin-coupled redox assay. It also has the capacity to reduce the organic compound tert-Butyl hydroperoxide (TBHP) in vitro, albeit at low efficiency. Sequence analysis, structural modeling, and site-directed mutagenesis revealed a conserved cysteine in the thioredoxin domain to be the redox active residue. Lastly, we provide evidence that recombinant human PhLP-3 exhibits redox activity similar to that of PbPhLP-3 and suggest that redox activity may be conserved in PhLP-3 homologs of other species. Our data provide new insight into the function of PhLP-3, which is hypothesized to act as co-chaperones in the folding and regulation of cytoskeletal proteins. We discuss the potential implications of PhLP-3 as a thioredoxin-target protein and possible links between the cellular redox network and the eukaryotic protein folding machinery.

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“…Since the discovery in which organoselenium compounds possess glutathione peroxidase (GPx) activity, a large number of the agent containing selenocysteines have been developed for different illnesses related to oxidative stress, such as cancer, Alzheimer’s, and inflammatory diseases. − In addition, some of these agents have also been utilized by immobilizing them on many substrates for the development of a surface enabling the generation of NO in a continuous manner. − Herein, the GPx activity of the synthesized catalyst, Di-DSePA, was evaluated using the nicotinamide adenine dinucleotide (NADPH)-reductase coupled assay. The assay measures a coupled reaction with a glutathione reductase (GR), in which oxidized glutathione catalyzed by GPx is reduced to glutathione in the presence of NADPH. , The oxidation of NADPH to NADP + corresponds to a decrease in the absorbance at 340 nm, where the rate of the decrease is directly proportional to the GPx activity of Di-DSePA …”

Section: Resultsmentioning

confidence: 99%

Endothelial Cell-Derived Tethered Lipid Bilayers Generating Nitric Oxide for Endovascular Implantation

Elnaggar

Hasan

Bhang

et al. 2021

ACS Appl. Bio Mater.

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Engineering an endothelium-mimetic surface has been one of long-lasting topics to develop ideal cardiovascular devices. The aim of the study was to investigate the potential use of a model of lipid bilayers that not only come from membranes extracted from endothelial cells (ECs) but also embedded with a type of organoselenium lipid enabling it to catalyze the generation of nitric oxide (NO). Herein, the titanium-cloaking in lipid bilayers extracted from ECs was prepared to propose a promising idea for the development of endovascular implants. For this purpose, we synthesized and characterized a lipidic molecule containing selenium and verified enough catalytic activity for the NO generation in the presence of S-nitrosothiols (RSNO) as endogenous NO precursors. We demonstrated the fabrication process of tethered lipid bilayers, from membrane extraction to vesicle fusion, and validated the successful formation of the layer and the catalyst insertion. The resulting bilayer presented endothelium-similar properties including the NO generation and cellular interactions. The catalyst inserted into the bilayer provided an unexampled result in the release period and kinetics of NO, likely similar to the native endothelial system. Using three different cells including EC, smooth muscle cell (SMC), and macrophage, it was demonstrated that the membrane responds selectively to each cell in the manner of promotive, suppressive, and nonimmunoreactive, respectively. Taken together, the fundamental study on obtained results not only provides understanding of the kinetics of designed NO catalyst and cellular interactions of reassembled membranes but also suggests very useful data on rational design and development of many vascular implantable devices, even expandable toward to nonvascular biointerfacing devices.

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Molecular cloning, characterization and expression profile of a glutathione peroxidase-like thioredoxin peroxidase (TPxGl) of the rodent malaria parasite Plasmodium berghei

Cited by 8 publications

References 36 publications

Functional genomic analyses of Enterobacter, Anopheles and Plasmodium reciprocal interactions that impact vector competence

Functional genomic analyses of Enterobacter, Anopheles and Plasmodium reciprocal interactions that impact vector competence

Characterization of a protozoan Phosducin-like protein-3 (PhLP-3) reveals conserved redox activity

Endothelial Cell-Derived Tethered Lipid Bilayers Generating Nitric Oxide for Endovascular Implantation

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