Identifying and characterising the Plasmodium falciparum RhopH3 Plasmodium vivax homologue

Mongui, Alvaro; Pérez-Leal, Oscar; Rojas-Caraballo, José; Angel, Diana I.; Cortés, Jimena; Patarroyo, Manuel A.

doi:10.1016/j.bbrc.2007.05.015

Cited by 18 publications

(23 citation statements)

References 31 publications

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“…A denaturing cycle was used which lasted 30 sec at 95°C followed by 35 cycles at 95°C for 10 sec, 58°C for 10 sec and 72°C for 2 min. The same conditions and previously described primers were used for amplifying the pvrhoph3 gene from cDNA [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

The Plasmodium vivax rhoptry neck protein 5 is expressed in the apical pole of Plasmodium vivax VCG-1 strain schizonts and binds to human reticulocytes

Arévalo-Pinzón

Bermúdez

Curtidor

et al. 2015

Malar J

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BackgroundDifferent proteins derived from the membrane or the apical organelles become involved in malarial parasite invasion of host cells. Among these, the rhoptry neck proteins (RONs) interact with a protein component of the micronemes to enable the formation of a strong bond which is crucial for the parasite’s successful invasion. The present study was aimed at identifying and characterizing the RON5 protein in Plasmodium vivax and evaluating its ability to bind to reticulocytes.MethodsTaking the Plasmodium falciparum and Plasmodium knowlesi RON5 amino acid sequences as template, an in-silico search was made in the P. vivax genome for identifying the orthologous gene. Different molecular tools were used for experimentally ascertaining pvron5 gene presence and transcription in P. vivax VCG-1 strain schizonts. Polyclonal antibodies against PvRON5 peptides were used for evaluating protein expression (by Western blot) and sub-cellular localization (by immunofluorescence). A 33 kDa PvRON5 fragment was expressed in Escherichia coli and used for evaluating the reactivity of sera from patients infected by P. vivax. Two assays were made for determining the RON5 recombinant fragment’s ability to bind to reticulocyte-enriched human umbilical cord samples.ResultsThe pvron5 gene (3,477 bp) was transcribed in VCG-1 strain schizonts and encoded a ~133 kDa protein which was expressed in the rhoptry neck of VCG-1 strain late schizonts, together with PvRON2 and PvRON4. Polyclonal sera against PvRON5 peptides specifically detected ~85 and ~30 kDa fragments in parasite lysate, thereby suggesting proteolytic processing in this protein. Comparative analysis of VCG-1 strain PvRON5 with other P. vivax strains having different geographic localizations suggested its low polymorphism regarding other malarial antigens. A recombinant fragment of the PvRON5 protein (rPvRON5) was recognized by sera from P. vivax-infected patients and bound to red blood cells, having a marked preference for human reticulocytes.ConclusionsThe pvron5 gene is transcribed in the VCG-1 strain, the encoded protein is expressed at the parasite’s apical pole and might be participating in merozoite invasion of host cells, taking into account its marked binding preference for human reticulocytes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0619-1) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

The Plasmodium vivax rhoptry neck protein 5 is expressed in the apical pole of Plasmodium vivax VCG-1 strain schizonts and binds to human reticulocytes

Arévalo-Pinzón

Bermúdez

Curtidor

et al. 2015

Malar J

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“…Peptide 37870 has been shown to be immunogenic in mice [44] and peptide 32416 has previously been used for polyclonal antibody production in rabbits, followed by localization experiments for the Pv RhopH3 protein [45]. Synthesized peptides were analysed by reverse phase high performance liquid chromatography (RP-HPLC) and MALDI-TOF mass spectrometry (Auoflex, Bruker Daltonics, Bremen, Germany).…”

Section: Methodsmentioning

confidence: 99%

Pv RON2, a new Plasmodium vivax rhoptry neck antigen

Arévalo-Pinzón

Curtidor

Patiño

et al. 2011

Malar J

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BackgroundRhoptries are specialized organelles from parasites belonging to the phylum Apicomplexa; they secrete their protein content during invasion of host target cells and are sorted into discrete subcompartments within rhoptry neck or bulb. This distribution is associated with these proteins' role in tight junction (TJ) and parasitophorous vacuole (PV) formation, respectively.MethodsPlasmodium falciparum RON2 amino acid sequence was used as bait for screening the codifying gene for the homologous protein in the Plasmodium vivax genome. Gene synteny, as well as identity and similarity values, were determined for ron2 and its flanking genes among P. falciparum, P. vivax and other malarial parasite genomes available at PlasmoDB and Sanger Institute databases. Pvron2 gene transcription was determined by RT-PCR of cDNA obtained from the P. vivax VCG-1 strain. Protein expression and localization were assessed by Western blot and immunofluorescence using polyclonal anti-PvRON2 antibodies. Co-localization was confirmed using antibodies directed towards specific microneme and rhoptry neck proteins.Results and discussionThe first P. vivax rhoptry neck protein (named here PvRON2) has been identified in this study. PvRON2 is a 2,204 residue-long protein encoded by a single 6,615 bp exon containing a hydrophobic signal sequence towards the amino-terminus, a transmembrane domain towards the carboxy-terminus and two coiled coil α-helical motifs; these are characteristic features of several previously described vaccine candidates against malaria. This protein also contains two tandem repeats within the interspecies variable sequence possibly involved in evading a host's immune system. PvRON2 is expressed in late schizonts and localized in rhoptry necks similar to what has been reported for PfRON2, which suggests its participation during target cell invasion.ConclusionsThe identification and partial characterization of the first P. vivax rhoptry neck protein are described in the present study. This protein is homologous to PfRON2 which has previously been shown to be associated with PfAMA-1, suggesting a similar role for PvRON2.

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“…To date, five P. vivax rhoptry proteins (RAP1 (Perez-Leal et al, 2006), RAP 2 (Patarroyo et al, 2005), RhopH3 (Mongui et al, 2007), Pv38 (Mongui et al, 2008), Pv34 (Mongui et al, 2009)) have been described by screening the complete P. vivax genome sequence (Carlton et al, 2008) and using a P. vivax strain adapted to Aotus monkeys as a source for parasite DNA, RNA and proteins (Pico de Coana et al, 2003). The characterized proteins might play an important role in human reticulocyte invasion and some of them have been evaluated as vaccine candidates in the Aotus animal model (Rojas-Caraballo et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Identifying and characterizing a member of the RhopH1/Clag family in Plasmodium vivax

Moreno-Pérez

Mongui

Soler

et al. 2011

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Identifying and characterising the Plasmodium falciparum RhopH3 Plasmodium vivax homologue

Cited by 18 publications

References 31 publications

The Plasmodium vivax rhoptry neck protein 5 is expressed in the apical pole of Plasmodium vivax VCG-1 strain schizonts and binds to human reticulocytes

The Plasmodium vivax rhoptry neck protein 5 is expressed in the apical pole of Plasmodium vivax VCG-1 strain schizonts and binds to human reticulocytes

Pv RON2, a new Plasmodium vivax rhoptry neck antigen

Identifying and characterizing a member of the RhopH1/Clag family in Plasmodium vivax

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