Molekulare Grundlage der Biogenese der Parasitophoren Vakuole von Plasmodium falciparum

顶复门是真核单细胞生物和专性细胞内寄生虫，其中医学上最相关的是恶性疟原虫。在侵入人类红细胞后，疟原虫启动膜的形成，其产生独特的隔室，寄生虫空泡，寄生虫驻留在宿主细胞内。从顶端分泌细胞器（如棒状体）分泌的蛋白质可能参与寄生虫液泡膜（PVM）的生物发生和维持。尽管PVM对寄生虫的细胞内生活起着关键作用，但PVM的大多数蛋白质组分尚未进行分子和功能表征。拟议项目的主要问题是分离PVM的分子库的细胞图谱蛋白质组学方法，使用二维聚丙烯酰胺凝胶电泳（2-D PAGE）结合质谱分析。来自与恶性疟原虫基因组计划相关的肽质量指纹的数据将允许识别PVM介导对寄生虫至关重要的功能的关键组分，例如将营养物质运输到液泡中和输出宿主细胞调节组分。动力学研究分析蛋白质的PVM疟原虫入侵和逃避之间的红细胞，重组表达选定的蛋白质的功能研究和免疫定位的合成细胞器将有助于阐明液泡膜的生物学作用。

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.