Molecular repertoire of rhoptry organelles from Plasmodium falciparum

棒状体是恶性疟原虫裂殖子中成对的分泌细胞器，并被认为在疟原虫红内期的宿主细胞入侵中起着至关重要的作用。关于顶端复合体的细胞器的功能，特别是棒状体，以及它们的关键组成部分的知识是稀缺的。拟议项目的主要问题是分析棒状体的分子库，即棒状体蛋白的鉴定和表征，其中大部分仍然是未知的。我们将通过差速离心富集棒状体细胞器，并通过双向凝胶电泳检测其蛋白质含量，双向凝胶电泳是一种已变得复杂且特别吸引人的生物体蛋白质组分析技术。在分离“棒状蛋白质组”后，将对几个突出的蛋白质点进行N-末端测序。新发现的蛋白质将被分子克隆以阐明其一级结构，并将序列与数据库中其他蛋白质的序列进行比较。重组表达选定的棒状体蛋白和产生针对这些因素的特异性抗体将有助于它们的功能特征，并阐明棒状体细胞器及其关键成分在寄生虫入侵中的作用。

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.