Development of a conditional localization approach to control apicoplast protein trafficking in malaria parasites.
Development of a conditional localization approach to control apicoplast protein trafficking in malaria parasites.
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开发一种条件定位方法来控制疟疾寄生虫中的顶质体蛋白运输。
DOI：
10.1111/tra.12656
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Prigge,SeanT
中科院分区：
文献类型：
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作者：
Roberts,AleahD;Nair,SethuC;Guerra,AlfredoJ;Prigge,SeanT
分泌蛋白对顶端复合体寄生虫特别重要，占导致疟疾、弓形虫病和巴贝斯虫病等疾病以及其他具有农业意义的疾病的人类病原体基因组的15%以上。在这里，我们开发了一种方法，使我们能够控制人类疟疾寄生虫恶性疟原虫中分泌蛋白的运输目的地。根据质外体转运肽的独特结构要求，我们设计了三个条件定位结构域(CLD1、2和3)，它们可以通过增加一个细胞配基来控制蛋白质的运输。比较每个CLD的贩运动态的研究表明，CLD2具有最优的贩运效率。为了验证这个系统，我们测试了CLD2是否可以在不干扰酶功能的情况下有条件地定位一种名为全羧基酶合成酶1(HCS1)的生物素连接酶。在一个表达CLD2-HCS1的寄生虫系中，我们能够以一种配体依赖的方式控制质外体中蛋白质的生物素化，证明了CLD工具的全部功能。我们已经开发并验证了一种新的分子工具，该工具可用于未来的研究，以帮助阐明疟疾寄生虫分泌蛋白的功能。
Secretory proteins are of particular importance to apicomplexan parasites and comprise over 15% of the genomes of the human pathogens that cause diseases like malaria, toxoplasmosis and babesiosis as well as other diseases of agricultural significance. Here, we developed an approach that allows us to control the trafficking destination of secretory proteins in the human malaria parasitePlasmodium falciparum. Based on the unique structural requirements of apicoplast transit peptides, we designed three conditional localization domains (CLD1, 2 and 3) that can be used to control protein trafficking via the addition of a cell permeant ligand. Studies comparing the trafficking dynamics of each CLD show that CLD2 has the most optimal trafficking efficiency. To validate this system, we tested whether CLD2 could conditionally localize a biotin ligase called holocarboxylase synthetase 1 (HCS1) without interfering with the function of the enzyme. In a parasite line expressing CLD2‐HCS1, we were able to control protein biotinylation in the apicoplast in a ligand‐dependent manner, demonstrating the full functionality of the CLD tool. We have developed and validated a novel molecular tool that may be used in future studies to help elucidate the function of secretory proteins in malaria parasites.