Stage-Specific and Selective Delivery of Caged Azidosugars into the Intracellular Parasite Toxoplasma gondii by Using an Esterase-Ester Pair Technique.
Stage-Specific and Selective Delivery of Caged Azidosugars into the Intracellular Parasite Toxoplasma gondii by Using an Esterase-Ester Pair Technique.
复制标题
使用酯酶-酯对技术将笼中的叠氮糖阶段特异性地选择性递送至细胞内寄生虫弓形虫中。
DOI：
10.1128/msphere.00142-19
复制
发表时间：
2019
期刊：
影响因子：
4.8
通讯作者：
Weiss,LouisM
中科院分区：
文献类型：
--
作者：
Tomita,Tadakimi;Wang,Hua;Wu,Peng;Weiss,LouisM
弓形虫是一种专性细胞内寄生虫，慢性感染多达三分之一的人口。寄生虫在中枢神经系统中以包囊的形式存在，并作为弓形虫脑炎再激活的宿主。已知囊壁具有丰富的O-连接的N-乙酰半乳糖胺聚糖，但是现有的代谢标记方法不允许在不标记宿主聚糖的情况下选择性标记胞内寄生虫糖蛋白。在这项研究中，我们整合了铜（I）催化的生物正交点击化学与特定的酯酶-酯对系统，以选择性地提供叠氮糖的细胞内寄生虫。我们证明α-环丙基修饰的GalNAz被寄生虫中产生的猪肝酯酶切割，而不是在宿主细胞中。我们的概念验证研究证明了这种酯酶-酯点击化学方法的可行性和潜力，用于以阶段特异性方式选择性地递送小分子。重要的是，由于存在多个膜和周围宿主细胞，选择性地将小分子递送到细胞内寄生虫中特别成问题。我们已经设计了一种方法，可以提供笼分子进入细胞内的寄生虫，弓形虫，表达uncaging酶在特定阶段的方式，而不影响宿主细胞的生物学。该系统为研究许多细胞内寄生虫提供了有价值的工具。
Toxoplasma gondii is an obligate intracellular parasite that chronically infects up to a third of the human population. The parasites persist in the form of cysts in the central nervous system and serve as a reservoir for the reactivation of toxoplasmic encephalitis. The cyst wall is known to have abundantO-linkedN-acetylgalactosamine glycans, but the existing metabolic labeling methods do not allow selective labeling of intracellular parasite glycoproteins without labeling of host glycans. In this study, we have integrated Cu(I)-catalyzed bioorthogonal click chemistry with a specific esterase-ester pair system in order to selectively deliver azidosugars to the intracellular parasites. We demonstrated that α-cyclopropyl modified GalNAz was cleaved by porcine liver esterase produced in the parasites but not in the host cells. Our proof-of-concept study demonstrates the feasibility and potential of this esterase-ester click chemistry approach for the selective delivery of small molecules in a stage-specific manner.IMPORTANCESelective delivery of small molecules into intracellular parasites is particularly problematic due to the presence of multiple membranes and surrounding host cells. We have devised a method that can deliver caged molecules into an intracellular parasite, Toxoplasma gondii, that express an uncaging enzyme in a stage-specific manner without affecting host cell biology. This system provides a valuable tool for studying many intracellular parasites.