Molecular repertoire of rhoptry organelles from Plasmodium falciparum

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Rhoptries are paired secretory organelles in Plasmodium falciparum merozoites and are suggested to play a crucial role in host cell invasion during erythrocytic stages of the malaria parasite. The knowledge about the function of the organelles of the apical complex, particularly of rhoptries, and about their key components is scarce. The major issue of the proposed project is the analysis of the molecular repertoire of rhoptries, i.e. the identification and characterization of rhoptry proteins, the majority of which is still unknown. We will enrich rhoptry organelles by differential centrifugationand examine their protein content by Two-dimensional gel electrophoresis, a technique which has become sophisticated andparticularly attractive for the analysis of proteoms of organisms. After separation of the "rhoptry proteom", severalprominent protein spots will be subjected to N-terminal sequencing. Newly identified proteins will be molecularly cloned to elucidate their primary structures, and the sequenceswill be compared with those of other proteins in data bases. Recombinant expression of selected rhoptry proteins and generation of specific antibodies against these factors will help to functionally characterize them and to elucidate the role of rhoptry organelles and their key components in parasiteinvasion.