Differential localization of processed fragments of Plasmodium falciparum serine repeat antigen and further processing of its N-terminal 47 kDa fragment.
The serine repeat antigen (SERA) of Plasmodium falciparum is a blood stage malaria vaccine candidate. It has been shown that 120 kDa SERA was proteolytically processed into N-terminal 47 kDa fragment (P47), central 56 kDa fragment (P56) that was further converted to 50 kDa (P50), and C-terminal 18 kDa fragment (P18). Here, we have examined the processing of SERA and the localization of its processed fragments by using mouse antibodies directed against recombinant proteins corresponding to different domains of SERA. Western blot analysis showed that all the processing events occurred inside parasitized erythrocytes at the stage just prior to the schizont rupture, that P47 was further processed into two 25 kDa fragments and that the two fragments, which were linked to P18 through disulfide bonds, were associated with the merozoite. In contrast, P50 was completely shed into culture medium and absent from the merozoite. This observation was further supported by the results of indirect immunofluorescence assay. These results could account for the findings that antibodies against P47 were inhibitory to the parasite growth in vitro but those against P50 were not. Finally, we demonstrated that the further processing of P47 is allelic type-dependent. The results of the present study would help in vaccine designing based on SERA.
Li
Jie
J
Department of Molecular Protozoology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
Mitamura
Toshihide
T
Fox
Barbara A
BA
Bzik
David J
DJ
Horii
Toshihiro
T
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology / Q000378:metabolism; D000953:Antigens, Protozoan / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D003470:Culture Media; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D021382:Protein Sorting Signals; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D013997:Time Factors
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003470:Culture Media; D010446:Peptide Fragments; D021382:Protein Sorting Signals; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C057495:serine repeat antigen, Plasmodium