A novel Plasmodium falciparum erythrocyte binding protein associated with the merozoite surface, PfDBLMSP.

Proteins on the surface of the merozoite, the invasive form of the malaria parasite Plasmodium falciparum,and those secreted from its apical secretory organelles are promising vaccine candidates against blood stage malaria. In the present study, we have identified a novel parasite protein (PfDBLMSP; Gene IDPF10_0348), that harbors a predicted signal sequence, a central Duffy binding-like (DBL) domain and a secreted polymorphic antigen associated with merozoites (SPAM) domain in its C-terminal half. Transcription and translation of pfdblmsp is up-regulated specifically in schizont stage parasites, similar to other well-chararacterized merozoite proteins involved in invasion of red blood cells (RBCs). PfDBLMSPwas localized on the merozoite surface with a GFP targeting approach using schizont-stage specific expression systems, and by immunofluorescence assays of the endogenous protein. PfDBLMSP expressed on the surface of mammalian cells (COS-7) showed binding with human RBCs and this binding was sensitive to trypsin and neuraminidase treatments. The recombinant proteins corresponding to the DBL and SPAM domains showed reactivity with immune sera from individuals residing in P. falciparum endemic areas. Polymorphism in PfDBLMSP sequences from different P. falciparum strains and field isolates suggested that its DBL domain is under natural immune pressure. Our data on localization and functional assays suggest a possible role of PfDBLMSP in binding of merozoites with erythrocytes during invasion.

Identification and characterization of a novel Plasmodium falciparum merozoite apical protein involved in erythrocyte binding and invasion.

Proteins that coat Plasmodium falciparum merozoite surface and those secreted from its apical secretory organelles are considered promising candidates for the vaccine against malaria. In the present study, we have identified an asparagine rich parasite protein (PfAARP; Gene ID PFD1105w), that harbors a predicted signal sequence, a C-terminal transmembrane region and whose transcription and translation patterns are similar to some well characterized merozoite surface/apical proteins. PfAARP was localized to the apical end of the merozoites by GFP-targeting approach using an inducible, schizont-stage expression system, by immunofluorescence assays using anti-PfAARP antibodies. Immuno-electron microsopic studies showed that PfAARP is localized in the apical ends of the rhoptries in the merozoites. RBC binding assays with PfAARP expressed on COS cells surface showed that it binds to RBCs through its N-terminal region with a receptor on the RBC surface that is sensitive to trypsin and neuraminidase treatments. Sequencing of PfAARP from different P. falciparum strains as well as field isolates showed that the N-terminal region is highly conserved. Recombinant protein corresponding to the N-terminal region of PfAARP (PfAARP-N) was produced in its functional form in E. coli. PfAARP-N showed reactivity with immune sera from individuals residing in P. falciparum endemic area. The anti-PfAARP-N rabbit antibodies significantly inhibited parasite invasion in vitro. Our data on localization, functional assays and invasion inhibition, suggest a role of PfAARP in erythrocyte binding and invasion by the merozoite.