An in silico down-scaling approach uncovers novel constituents of the Plasmodium-containing vacuole

Matz, Joachim M.; Matuschewski, Kai

doi:10.1038/s41598-018-32471-6

Cited by 9 publications

(21 citation statements)

References 43 publications

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“…The PV and FV localization of PfLCN are in agreement with a recent study, in which the PfLCN orthologue in the rodent malaria model P. berghei (termed PV5) was identified as an essential PV protein (Matz & Matuschewski, 2018). The FV localization is further supported by mass spectrometric analysis, which showed that PfLCN is part of the FV proteome (Lamarque et al, 2008).…”

Section: Discussionsupporting

confidence: 90%

Structure-based identification and functional characterization of an essential lipocalin in the malaria parasitePlasmodium falciparum

Burda

Crosskey

Lauk

et al. 2020

Preprint

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Proteins of the lipocalin family are known to bind small hydrophobic ligands and are involved in various physiological processes ranging from lipid transport to oxidative stress responses. The genome of the malaria parasite Plasmodium falciparum contains a single protein PF3D7_0925900 with a lipocalin signature. Using crystallography and small-angle X-ray scattering, we show that the protein has a tetrameric structure of typical lipocalin monomers, hence we name it P. falciparum lipocalin (PfLCN), the first lipocalin structurally and functionally characterized in a single-celled eukaryote. We show that PfLCN is expressed in the intraerythrocytic stages of the parasite and localizes to the parasitophorous and food vacuoles. Conditional knockdown of PfLCN impairs parasite development, which can be rescued by treatment with the radical scavenger Trolox or by temporal inhibition of hemoglobin digestion. This suggests a key function of PfLCN in counteracting oxidative stress induced cell damage during multiplication of parasites within red blood cells.Keywords: blood stage / food vacuole / lipocalin / malaria / oxidative stress compound 2. Images and time-lapse movies were acquired on microscopes of the CSSB imaging facility.

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Section: Discussionsupporting

confidence: 90%

Structure-based identification and functional characterization of an essential lipocalin in the malaria parasitePlasmodium falciparum

Burda

Crosskey

Lauk

et al. 2020

Preprint

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“…Live fluorescence microscopy of transgenic P. berghei parasites expressing mCherry-3xMyc-tagged Pb PV5 confirmed that the protein localizes to tubular PV extensions during ring and trophozoite stages and surrounds individual merozoites in segmented schizonts (Fig. 1 C ) (27). In addition, a substantial fraction of the protein was restricted to the parasite cytoplasm.…”

Section: Resultsmentioning

confidence: 82%

“…Employing a genome-wide in silico down-scaling approach, we previously identified an essential P. berghei PV protein, Pb PV5 (PBANKA_0826700), which has orthologues in all other Plasmodium species, including P. falciparum (PF3D7_0925900) (27). Inspection of the PV5 amino acid sequence revealed a striking similarity to members of the functionally diverse lipocalin family, barrel-shaped proteins capable of binding various hydrophobic ligands and/or protein interaction partners (28).…”

Section: Resultsmentioning

confidence: 99%

“…( C ) Dual protein localization of PV5 to extensions of the PV and to intraparasitic structures in P. berghei . Transgenic parasites expressing the PV marker GFP PV and the endogenous Pb PV5 gene fused to mCherry-3xMyc were imaged live (27). Shown are the mCherry (red, first row) and GFP channels (green, second row), a merge of both signals (third row) and a merge of differential interference contrast images (DIC) and Hoechst 33342 nuclear stain (blue, fourth row).…”

Section: Resultsmentioning

confidence: 99%

“…Our previous attempts to disrupt the genomic Pb PV5 locus resulted only in atypical integration of the targeting construct without perturbing the endogenous gene, which is indicative of essential functions during the asexual blood stage cycle in vivo (27). As an alternative genetic strategy to analyse Pb PV5 function, we sought to deregulate Pb PV5 expression by employing a promoter swap approach (Fig.…”

Section: Resultsmentioning

confidence: 99%

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A lipocalin mediates unidirectional haem biomineralization in malaria parasites

Matz

Drepper

Blum

et al. 2020

Preprint

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18During blood stage development, malaria parasites are challenged with the detoxification of 19 enormous amounts of haem released during the proteolytic catabolism of erythrocytic 20 haemoglobin. They tackle this problem by sequestering haem into bioinert crystals known as 21 haemozoin. The mechanisms underlying this biomineralization process remain enigmatic. 22Here, we demonstrate that both rodent and human malaria parasite species secrete and 23 internalize a lipocalin-like protein, PV5, to control haem crystallization. Transcriptional 24 deregulation of PV5 in the rodent parasite Plasmodium berghei results in inordinate elongation 25 of haemozoin crystals, while conditional PV5 inactivation in the human malaria agent 26 Plasmodium falciparum causes excessive multi-directional crystal branching. Although 27 haemoglobin processing remains unaffected, PV5-deficient parasites generate less 28 haemozoin. Electron diffraction analysis indicates that despite the distinct changes in crystal 29 morphology neither the crystalline order nor unit cell of haemozoin are affected by impaired 30 PV5 function. Deregulation of PV5 expression renders P. berghei hypersensitive to the 31 antimalarial drugs artesunate, chloroquine, and atovaquone, resulting in accelerated parasite 32 clearance following drug treatment in vivo. Together, our findings demonstrate the 33 Plasmodium-tailored role of a lipocalin family member in haemozoin formation and underscore 34 the haem biomineralization pathway as an attractive target for therapeutic exploitation.35 SIGNIFICANCE 37During blood stage development, the malaria parasite replicates inside erythrocytes of the 38 vertebrate host, where it engulfs and digests most of the available haemoglobin. This results 39 in release of the oxygen-binding prosthetic group haem, which is highly toxic in its unbound 40 form. The parasite crystallizes the haem into an insoluble pigment called haemozoin, a 41 process that is vital for parasite survival and which is exploited in antimalarial therapy. We 42 demonstrate that the parasite uses a protein called PV5 in haemozoin formation and that 43 interfering with PV5 expression can increase the parasite's sensitivity to antimalarial drugs 44 during blood infection. An improved understanding of the mechanisms underlying haem 45 sequestration will provide valuable insights for future drug development efforts.(1). Throughout blood stage development, the parasite ingests and catabolizes up to 80% of 50 the host cell cytoplasm, facilitating amino acid acquisition and making sufficient room for 51 parasite growth (2, 3). Haemoglobin is incorporated through endocytosis and then degraded 52 by an array of functionally redundant proteases, a process which occurs in acidified lysosome-53 like organelles with species-specific morphology (4). In the rodent-infective parasite species 54 Plasmodium berghei, one or more food vacuoles (FVs) give rise to small digestive vesicles 55 (DVs) which only fuse at the very end of intraerythrocytic development (5). By contrast, the...

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Structure-Based Identification and Functional Characterization of a Lipocalin in the Malaria Parasite Plasmodium falciparum

et al. 2020

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An in silico down-scaling approach uncovers novel constituents of the Plasmodium-containing vacuole

Cited by 9 publications

References 43 publications

Structure-based identification and functional characterization of an essential lipocalin in the malaria parasitePlasmodium falciparum

Structure-based identification and functional characterization of an essential lipocalin in the malaria parasitePlasmodium falciparum

A lipocalin mediates unidirectional haem biomineralization in malaria parasites

Structure-Based Identification and Functional Characterization of a Lipocalin in the Malaria Parasite Plasmodium falciparum

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