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 parasite Plasmodium 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.

中文翻译：

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有条件的本地化方法的发展，以控制疟疾寄生虫中的原生质膜蛋白运输。

分泌蛋白对蚜虫寄生虫特别重要，并且占导致引起诸如疟疾，弓形虫病和巴贝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。基于apicoplast运输肽的独特结构要求，我们设计了三个条件定位域（CLD1、2和3），可通过添加细胞渗透性配体来控制蛋白质运输。比较每个CLD的交易动态的研究表明，CLD2具有最佳的交易效率。要验证此系统，我们测试了CLD2是否可以有条件地定位称为全羧化酶合成酶1（HCS1）的生物素连接酶，而不会干扰该酶的功能。在表达CLD2-HCS1的寄生虫品系中，我们能够以配体依赖性方式控制无纺布中的蛋白质生物素化，这证明了CLD工具的全部功能。我们已经开发并验证了一种新颖的分子工具，该工具可用于将来的研究中，以帮助阐明疟原虫中分泌蛋白的功能。