Reconstitution of Plasmodium Export in Toxoplasma

弓形虫中疟原虫输出的重建

基本信息

批准号：
8463994
负责人：
Peter John Bradley
金额：
$ 7.32万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8463994
关键词：
ATP phosphohydrolase Architecture Aspartic Endopeptidases Binding Biological Assay Cells Chimeric Proteins Codon Nucleotides Complex Cytoplasm Data Development Disease Elements Endoplasmic Reticulum Engineering Erythrocytes Funding Mechanisms Host Defense Human Immunocompromised Host Invaded Ion Channel Licensing Life Style Malaria Mediating Medical Membrane Names Nutrient Parasites Pathway interactions Peptide Signal Sequences Plasmodium Plasmodium falciparum Plasmodium malariae Process Property Protein Export Pathway Protein translocation Proteins Relative (related person)Reporter System Testing Therapeutic Intervention Toxin Toxoplasma Toxoplasma gondii Toxoplasmosis Transmembrane Transport Transport Process Vacuole Virulence cell type design neonate new technology novel novel strategies obligate intracellular parasite pathogen plasmepsin protein complex protein transport reconstitution secretory protein technology development thioredoxin-like protein

项目摘要

DESCRIPTION (provided by applicant): Apicomplexan parasites such as Plasmodium falciparum and Toxoplasma gondii share a common obligate intracellular lifestyle in which the parasite actively penetrates the host cell and resides in a unique membrane-bound vacuole in the cytoplasm of the host. While these parasites are sequestered inside the vacuole, an emerging paradigm is that apicomplexans also deliver proteins into the host cell to modulate the host for optimal intracellular survival. This is particularly important in the intraerythrocytic stges of the Plasmodia, as the red blood cell host is largely metabolically inactive and devoid of many membrane transport systems and organellar functions that can be co-opted in other cells. To compensate for the relative lack of host functions to hijack, Plasmodium efficiently exports hundreds of secretory proteins across the vacuolar membrane and into the host cell that dramatically remodel of the erythrocyte and are key regulators of parasite virulence. Protein export occurs by a specialized vacuolar membrane transport apparatus known as the PTEX translocon, which is unique to Plasmodium and essential for parasite survival. In this proposal, we exploit the similar properties of the secretory pathway and parasitophorous vacuole in apicomplexans to reconstitute the PTEX translocon and export pathway in Toxoplasma. As an initial proof of concept, we have expressed two key components of the translocon, EXP2 and HSP101, in T. gondii and shown that they are secreted to the parasitophorous vacuole membrane. We will build on these results by expressing the remaining three translocon components and assessing their localization and ability to form a complex at the vacuolar membrane. We will also express a Plasmodium host-targeted reporter protein to determine if we can establish a functional PTEX-mediated export pathway in T. gondii. Development of this technology will enable new approaches to dissect the assembly, architecture and function of the PTEX translocon, and also enable the design of novel therapies that specifically target this critical protein export pathway.

描述（由申请人提供）：apicomplexan寄生虫，例如恶性疟原虫和弓形虫弓形虫，具有共同的义务性义务细胞内生活方式，在这种生活方式中，寄生虫在宿主的细胞质中积极渗透宿主细胞并驻留在独特的膜结合液泡中。虽然这些寄生虫在液泡内部隔离，但新兴的范式是，Apicomplexans还将蛋白质输送到宿主细胞中以调节宿主以获得最佳的细胞内存活。这在疟原虫的肉体内stges中尤为重要，因为红细胞宿主在很大程度上是代谢不活跃的，并且没有许多可以在其他细胞中选择的膜转运系统和细胞器功能。为了弥补劫持剂的相对缺乏宿主功能，疟原虫有效地将数百种分泌蛋白在整个液泡膜上出口到宿主细胞中，这些蛋白会急剧重塑红细胞的重塑，并且是寄生虫病毒性的关键调节剂。蛋白质导出是由一种专门的液泡膜转运设备称为PTEX易位，该设备是疟原虫独有的，对于寄生虫生存所必需。在此提案中，我们利用了Apicomplexans中分泌途径和寄生虫液泡的相似特性来重建弓形虫中的PTEX Clressocon和Export Pathway。作为最初的概念证明，我们在T. gondii中表达了Clressocon，Exp2和Hsp101的两个关键组成部分，并表明它们被分泌到寄生虫液泡膜上。我们将通过表达剩余的三个转运成分并评估其本地化和在液泡膜上形成复合物的能力来建立这些结果。我们还将表达疟原虫靶向的报告蛋白，以确定是否可以在T. gondii中建立功能性PTEX介导的导出途径。这项技术的开发将使新方法能够剖析PTEX Clressocon的组装，结构和功能，并能够设计出专门针对这种关键蛋白质导出途径的新型疗法。