Sugar nucleotide quantification by liquid chromatography tandem mass spectrometry reveals a distinct profile in Plasmodium falciparum sexual stage parasites.
Sugar nucleotide quantification by liquid chromatography tandem mass spectrometry reveals a distinct profile in Plasmodium falciparum sexual stage parasites.
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DOI：
10.1042/bcj20161030
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发表时间：
2017-03-07
期刊：
影响因子：
--
通讯作者：
Izquierdo L
中科院分区：
文献类型：
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作者：
López-Gutiérrez B;Dinglasan RR;Izquierdo L
恶性疟原虫的专性细胞内生活方式和难以获得足够量的生物材料，阻碍了疟疾寄生虫的特定代谢途径的研究。因此，例如，燃料糖基化反应所需的糖核苷酸库从未在同步良好的无性寄生虫或其生命周期的其他阶段进行过深入研究。这些代谢物至关重要，特别是考虑到对关键寄生虫蛋白中存在的N-、O-和其他聚糖的重新关注。在这项工作中，我们采用了液相色谱串联质谱（LC-MS/MS）方法的基础上使用多孔石墨碳（PGC）柱和MS友好的溶剂来量化疟疾寄生虫中的糖核苷酸。我们报告了在恶性疟原虫的红细胞内循环中这些代谢物池的彻底定量。该方法的灵敏度首次实现了对配子体中这些糖基化前体的靶向分析，配子体是可传播给蚊子载体的寄生虫性阶段。
The obligate intracellular lifestyle of Plasmodium falciparum and the difficulties in obtaining sufficient amounts of biological material have hampered the study of specific metabolic pathways in the malaria parasite. Thus, for example, the pools of sugar nucleotides required to fuel glycosylation reactions have never been studied in-depth in well-synchronized asexual parasites or in other stages of its life cycle. These metabolites are of critical importance, especially considering the renewed interest in the presence of N-, O-, and other glycans in key parasite proteins. In this work, we adapted a liquid chromatography tandem mass spectrometry (LC-MS/MS) method based on the use of porous graphitic carbon (PGC) columns and MS-friendly solvents to quantify sugar nucleotides in the malaria parasite. We report the thorough quantification of the pools of these metabolites throughout the intraerythrocytic cycle of P. falciparum. The sensitivity of the method enabled, for the first time, the targeted analysis of these glycosylation precursors in gametocytes, the parasite sexual stages that are transmissible to the mosquito vector.