Molekulare Grundlage der Biogenese der Parasitophoren Vakuole von Plasmodium falciparum

顶体复合体是真核单细胞生物，是细胞内的必备寄生虫，在医学上与恶性疟原虫关系最密切。当疟原虫侵入人类红细胞时，它会启动一层膜的形成，从而形成一个独特的隔间--寄生虫液泡，寄生虫就住在这个液泡里。顶端分泌细胞器释放的蛋白质可能参与寄生性液泡膜(PVM)的生物发生和维持。尽管PVM对寄生虫S的细胞内生命起着关键作用，但PVM的大部分蛋白质成分还没有得到分子和功能的表征。该项目的主要问题是利用双向聚丙烯酰胺凝胶电泳法(2-D PAGE)和质谱仪相结合的细胞图谱蛋白质组学方法分析分离的PVM的分子谱带。来自与恶性疟原虫基因组计划相关的多肽质量指纹的数据将使我们能够识别PVM的关键成分，这些成分对寄生虫至关重要的中介功能，如将营养物质运输到液泡和输出宿主细胞调节成分。对PVM蛋白在原虫侵入和逃逸红细胞之间的动力学分析、功能研究中选择的重组蛋白的表达以及它们对合成细胞器的免疫定位将有助于阐明空泡膜的生物学作用。

Apicomplexa are eukaryotic unicellular organisms and obligatory intracellular parasites, the medically most relevant of which is Plasmodium falciparum. Upon invasion into human erythrocytes, Plasmodium initiates the formation of a membrane which creates a unique compartment, the parasitophorous vacuole, in which the parasite resides inside the host cell. Proteins discharged from apical secretory organelles, e.g. the rhoptries, are presumably involved in the biogenesis and maintenance of the parasitophorous vacuole membrane (PVM). Despite the pivotal role of the PVM for the parasite´s intracellular life, most of the protein components of the PVM have not been molecularly and functionally characterized. The major issue of the proposed project is the analysis of the molecular repertoire of isolated PVM by a cell-map proteomic approach using two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) combined with mass spectrometry. Data from peptide mass finger prints linked to the genome project of P. falciparum will allow to identify key components of the PVM mediating functions vital for the parasite such as transport of nutrients into the vacuole and export of host cell modulating components. Kinetic studies analyzing the proteins of the PVM between plasmodial invasion into and evasion from the red blood cell, recombinant expression of selected proteins for functional studies and their immunolocalization to the synthesizing organelles will help to elucidate the biological role of the vacuolar membrane.