Identification of an endoplasmic reticulum-resident calcium-binding protein with multiple EF-hand motifs in asexual stages of Plasmodium falciparum1Note: Nucleotide sequence data reported in this paper have been deposited in the GenBank™ data base with the accession number AF016410.1

Greca, Nadia La; Hibbs, Alan R.; Riffkin, Christopher D.; Foley, Michael; Tilley, Leann

doi:10.1016/s0166-6851(97)00134-5

Cited by 75 publications

(23 citation statements)

References 44 publications

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“…To investigate the function of Plasmepsin V, we introduced a hemagglutinin (HA) tag at the C-terminus (3D7-PMVHA) (Fig 2a). HA-tagged Plasmepsin V localised to the ER 25, as has been shown previously for the endogenous protein 24 (Fig. 2b).…”

Section: Plasmepsin V Is An Er Resident Aspartic Acid Protease That Csupporting

confidence: 83%

“…Cells were incubated with rabbit α-Pf ERC (1:500) 25, 3F10 rat α-HA (1:50) or mouse α-Plasmepsin V (1:5) for 2 hr followed by Alexa Fluor 488- or 594-conjugated secondary IgG antibodies (1:1000; Molecular Probes) for 1 hr and nuclei were labelled with DAPI nuclear stain (Roche) at 0.2 μg/ml in Vectorshield (Vector Labs). Cells were viewed with a Carl Zeiss Axioskop 2 microscope (Thornwood) and images collected using a PCO SensiCam (Motion Engineering Co.) and Axiovision 3 software (Carl Zeiss).…”

Section: Methodsmentioning

confidence: 99%

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An aspartyl protease directs malaria effector proteins to the host cell

Boddey

Hodder

Günther

et al. 2010

Nature

298

417

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Plasmodium falciparum causes the virulent form of malaria and disease manifestations are linked to growth inside infected erythrocytes. In order to survive and evade host responses the parasite remodels the erythrocyte by exporting several hundred effector proteins beyond the surrounding parasitophorous vacuole membrane. A feature of exported proteins is a pentameric motif (RxLxE/Q/D) that is a substrate for an unknown protease. Here, we show the protein responsible for cleavage of this motif is Plasmepsin V, an aspartic acid protease located in the endoplasmic reticulum. Plasmepsin V cleavage reveals the export signal (xE/Q/D) at the N-terminus of cargo proteins. Expression of an identical mature protein with xQ at the N-terminus generated by signal peptidase was not exported demonstrating Plasmepsin V activity is essential and linked with other key export events. Identification of the protease responsible for export into erythrocytes provides a novel target for therapeutic intervention against this devastating disease.

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Section: Plasmepsin V Is An Er Resident Aspartic Acid Protease That Csupporting

confidence: 83%

Section: Methodsmentioning

confidence: 99%

An aspartyl protease directs malaria effector proteins to the host cell

Boddey

Hodder

Günther

et al. 2010

Nature

298

417

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“…Treatment of the transfected parasitized erythrocytes with 4 or 8 hemolytic units of SLO released more than 97% of the hemoglobin from the parasitized erythrocytes (data not shown), but did not release PfERC (Fig. 4C), which has previously been shown to be located within the ER lumen (34). S-antigen (a PV marker (35)) was also largely associated with the pellet fraction (0.5 and 4% in the supernatant, respectively, for 4 and 8 units of SLO; Fig.…”

Section: Exp1-(1-35) Is Present In the Pv And Fv Of Transfected P Famentioning

confidence: 63%

The Signal Sequence of Exported Protein-1 Directs the Green Fluorescent Protein to the Parasitophorous Vacuole of Transfected Malaria Parasites

Adisa¹,

Rug²,

Klonis

et al. 2003

Journal of Biological Chemistry

Self Cite

114

108

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The malaria parasite, Plasmodium falciparum, spends part of its life cycle inside the erythrocytes of its human host. In the mature stages of intraerythrocytic growth, the parasite undertakes extensive remodeling of its adopted cellular home by exporting proteins beyond the confines of its own plasma membrane. To examine the signals involved in export of parasite proteins, we have prepared transfected parasites expressing a chimeric protein comprising the N-terminal region of the Plasmodium falciparum exported protein-1 appended to green fluorescent protein. The majority of the population of the chimeric protein appears to be correctly processed and trafficked to the parasitophorous vacuole, indicating that this is the default destination for protein secretion. Some of the protein is redirected to the parasite food vacuole and further degraded. Photobleaching studies reveal that the parasitophorous vacuole contains subcompartments that are only partially interconnected. Dual labeling with the lipid probe, BODIPY-TR-ceramide, reveals the presence of membrane-bound extensions that can bleb from the parasitophorous vacuole to produce double membranebound compartments. We also observed regions and extensions of the parasitophorous vacuole, where there is segregation of the lumenal chimera from the lipid components. These regions may represent sites for the sorting of proteins destined for the trafficking to sites beyond the parasitophorous vacuole membrane.Due to the compartmentalization of eukaryotic cells, a sophisticated protein trafficking system is an integral requirement for homeostasis and growth. Proteins destined for compartments other than the cytoplasm are synthesized with intrinsic signals that determine their transport within the cell. Small peptide motifs often form the necessary targeting determinants (1). For example, an N-terminal hydrophobic sequence forms part of the typical secretory signal that directs proteins across the endoplasmic reticulum (ER) 1 membrane (2-5). Similarly, N-terminal amphipathic and bipartite sequences target proteins to the chloroplast and mitochondria (6 -8).The malaria parasite, Plasmodium falciparum, spends part of its life cycle inside mature human erythrocytes. The parasite invades this quiescent host cell and develops within a parasitophorous vacuole (PV). An unusual and highly specialized secretory system enables the malaria parasite to survive within a cell that lacks its own machinery for protein synthesis and trafficking. Indeed, the parasite targets proteins, not only to compartments within its own confines, but to the PV, in which it resides, as well as the PV membrane (PVM), the erythrocyte cytoplasm, and host cell membrane (9 -11).Efforts have been made to understand the trafficking signals that target parasite proteins to different compartments within and outside the parasite. Proteins destined for the ER, the parasite plasma membrane (PPM), the PV, or the PVM appear to have a "classical" hydrophobic N-terminal signal sequence (i.e. a stretch of 10 -15 hydrop...

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“…Some gene sequences potentially encode functional CBLs in T. gondii and P. falciparum . A reticulocalbin located in the ER of asexual stages of P. falciparum contains 6 EF-hand domains [61]. …”

Section: Calcium Binding Proteins In Apicomplexan Parasitesmentioning

confidence: 99%

The calcium signaling toolkit of the Apicomplexan parasites Toxoplasma gondii and Plasmodium spp

2015

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Apicomplexan parasites have complex life cycles, frequently split between different hosts and reliant on rapid responses as the parasites react to changing environmental conditions. Calcium ion (Ca2+) signaling is consequently essential for the cellular and developmental changes that support apicomplexan parasitism. Apicomplexan genomes reveal a rich repertoire of genes involved in calcium signaling, although many of the genes responsible for observed physiological changes remain unknown. There is evidence, for example, for the presence of a nifedipine-sensitive calcium entry mechanism in Toxoplasma, but the molecular components involved in Ca2+ entry in both Toxoplasma and Plasmodium, have not been identified. The major calcium stores are the endoplasmic reticulum (ER), the acidocalcisomes, and the plant-like vacuole in Toxoplasma, or the food vacuole in Plasmodium spp. Pharmacological evidence suggests that Ca2+ release from intracellular stores may be mediated by inositol 1,4,5-trisphosphate (IP3) or cyclic ADP ribose (cADPR) although there is no molecular evidence for the presence of receptors for these second messengers in the parasites. Several Ca2+-ATPases are present in apicomplexans and a putative mitochondrial Ca2+/H+ exchanger has been identified. Apicomplexan genomes contain numerous genes encoding Ca2+-binding proteins, with the notable expansion of calcium-dependent protein kinases (CDPKs), whose study has revealed novel roles in gliding motility, microneme secretion, host cell invasion and egress, and parasite differentiation. Microneme secretion has also been shown to depend on the C2 domain containing protein DOC2 in both Plasmodium spp. and Toxoplasma, providing further evidence for the complex transduction of Ca2+ signals in these organisms. The characterization of these pathways could lead to the discovery of novel drug targets and to a better understanding of the role of Ca2+ in these parasites.

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Identification of an endoplasmic reticulum-resident calcium-binding protein with multiple EF-hand motifs in asexual stages of Plasmodium falciparum1Note: Nucleotide sequence data reported in this paper have been deposited in the GenBank™ data base with the accession number AF016410.1

Cited by 75 publications

References 44 publications

An aspartyl protease directs malaria effector proteins to the host cell

An aspartyl protease directs malaria effector proteins to the host cell

The Signal Sequence of Exported Protein-1 Directs the Green Fluorescent Protein to the Parasitophorous Vacuole of Transfected Malaria Parasites

The calcium signaling toolkit of the Apicomplexan parasites Toxoplasma gondii and Plasmodium spp

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