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[Fe-S] cluster assembly in the apicoplast and its indispensability in mosquito stages of the malaria parasite

机译:[Fe-S]聚集在疟疾寄生虫蚊子阶段的蚊子阶段的不可或缺

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摘要

The relict plastid (apicoplast) of the malaria parasite is the site for important biochemical pathways and is essential for parasite survival. The sulfur mobilization (SUF) pathway of iron-sulfur [Fe-S] cluster assembly in the apicoplast of Plasmodium spp. is of interest due to its absence in the human host suggesting the possibility of antimalarial intervention through apicoplast [Fe-S] biogenesis. We report biochemical characterization of components of the Plasmodium falciparum apicoplast SUF pathway after the first step of SUF. In vitro interaction experiments and in vivo cross-linking showed that apicoplast-encoded PfSufB and apicoplast-targeted PfSufC and PfSufD formed a complex. The PfSufB-C2-D complex could function as a scaffold to assemble [4Fe-4S] clusters in vitro and activity of the PfSufC ATPase was enhanced by PfSufD. Two carrier proteins, the NifU-like protein PfNfu and the A-type carrier PfSufA are homodimers, the former mediating transfer of [4Fe-4S] from the scaffold to a model [4Fe-4S] target protein with higher efficiency. Conditional knockout of SufS, the enzyme catalyzing the first step of SUF, by selective excision in the mosquito stages of Plasmodium berghei severely impaired development of sporozoites in oocysts establishing essentiality of the SUF machinery in the vector. Our results delineate steps of the complete apicoplast SUF pathway and demonstrate its critical role in the parasite life cycle.
机译:疟疾寄生虫的relict plastid(apicoplast)是重要的生化途径的部位,对于寄生虫生存至关重要。在疟原虫SPP的支机构中的铁 - 硫[Fe-S]簇组装的硫动力(SUF)途径。由于其在人宿主中的缺失,表明通过apicoplast [Fe-S]生物发生抗疟疾干预的可能性。我们报告了SUF的第一步之后的疟原虫疟原虫Apicoplast SUF途径组分的生化表征。体外相互作用实验和体内交联显示出Apicoplast编码的PFSUFB和Apicoplast靶向PFSUFC和PFSUFD形成了复合物。 PFSUFB-C2-D复合物可以用作支架以组装[4FE-4S]体外体外簇,PFSUFD增强了PFSUFC ATP酶的活性。两种载体蛋白质,NiFu样蛋白PFNFU和A型载体PFSUFA是同型二聚体,前者将[4FE-4S]从支架中介导的[4FE-4S]靶蛋白的效率高。 Sufs的条件敲除,催化SUF第一步的酶,通过在Plasmodium Perghei的蚊子阶段选择性切除严重受损地损害了卵囊中的孢子生殖物的发育,在载体中建立了SUF机械的本质。我们的结果描绘了完整的Apicoplast SUF途径的步骤,并在寄生虫生命周期中展示其关键作用。

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