Plasmodium falciparum rhoptry proteins of 140/130/110 kd (Rhop-H) are located in an electron lucent compartment in the neck of the rhoptries.
To investigate in more detail the structure of the high molecular weight rhoptry protein complex of Plasmodium falciparum, Rhop-H (140/130/110 kd), the complex was affinity purified from parasite extracts using rhoptry protein specific antisera prepared against Rhop-H proteins bound to and eluted from Balb/c mouse erythrocytes, using 0.5 M NaCl. The individual proteins (140 kd/Rhop-1, 130 kd/Rhop-2, and 110 kd/Rhop-3) were separated, electroeluted, and monospecific polyclonal antisera prepared against the individual proteins, and against the affinity purified complex. Immunofluorescence assays and immunoelectron microscopic studies were performed to verify the subcellular localization of the Rhop-H epitopes. Immunoblotting and immunoprecipitation assays were also performed. We report novel findings regarding the localization of the rhoptry proteins to an electron lucent compartment in the neck of the rhoptries. Analysis of the amino acid composition of the individually purified Rhop-H proteins demonstrated a predominance of negatively charged (E, D) as well as hydrophobic residues (L, A, P, S) in the three proteins. The percentage of negatively charged residues was high for all three proteins. Similarities in amino acid composition for the three proteins supports the previous data demonstrating shared properties such as erythrocyte and liposome binding, for the three proteins. Results of antibody characterizations using rhoptry protein specific antisera demonstrate the immunodominance of the Rhop-H complex.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000918:Antibody Specificity; D000953:Antigens, Protozoan / Q000032:analysis*; D002846:Chromatography, Affinity; D004912:Erythrocytes / Q000469:parasitology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007074:Immunoglobulin G; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010963:Plasmodium falciparum / Q000469:parasitology / Q000502:physiology* / Q000648:ultrastructure; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D011817:Rabbits / Q000276:immunology
D000596:Amino Acids; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D010446:Peptide Fragments; D015800:Protozoan Proteins; C065352:rhoptry associated protein, Plasmodium