CE-ESI-MS metabolic fingerprinting of Leishmania resistance to antimony treatment
CE-ESI-MS metabolic fingerprinting of Leishmania resistance to antimony treatment
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DOI：
10.1002/elps.201200007
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发表时间：
2012-07-01
期刊：
影响因子：
2.9
通讯作者：
Barbas, Coral
中科院分区：
文献类型：
--
作者：
Canuto, Gisele A. B.;Castilho-Martins, Emerson A.;Barbas, Coral
代谢组学已成为揭示病原体生物学的宝贵工具，并立即应用于化疗。目前公认的是，没有一种单一的技术能够获得生物系统的整个代谢指纹，无论是由于它们的不同的物理化学性质或浓度。在这项工作中，我们已经探索了毛细管电泳质谱与无鞘接口与电喷雾电离（CE-ESI-TOF-MS）分离代谢物，以作为一种补充技术，以LC的能力。作为概念的证明，我们比较了婴儿利什曼原虫前鞭毛体BCN 150（Sb（III）IC 50 = 20.9 μ M）的代谢组和其变化时，用120 μ M的Sb（III）酒石酸钾处理12小时，以及与Sb（III）的抗性对应物，通过逐步增加Sb（III）下的寄生虫的生长获得高达180 μ M。BCN 150 Sb（III）处理与未处理的代谢产物数量比较为264，Sb（III）抗性与敏感寄生虫的代谢产物数量比较为195。在连续的数据过滤后，利什曼原虫途径数据库中鉴定的7种代谢物的差异显示出最高的显著性差异，主要对应于氨基酸或其代谢物替代物。他们中的大多数被分配到含硫氨基酸和多胺的生物合成途径，考虑到锑（III）的利什曼原虫的巯基依赖的氧化还原代谢的恶化特别相关。考虑到大多数代谢物的典型低浓度，可以认为该试验是成功的，应探索新的生物学问题。
Metabolomics has become an invaluable tool to unveil biology of pathogens, with immediate application to chemotherapy. It is currently accepted that there is not one single technique capable of obtaining the whole metabolic fingerprint of a biological system either due to their different physical-chemical properties or concentrations. In this work, we have explored the capability of capillary electrophoresis mass spectrometry with a sheathless interface with electrospray ionization (CE-ESI-TOF-MS) to separate metabolites in order to be used as a complementary technique to LC. As proof of concept, we have compared the metabolome of Leishmania infantum promastigotes BCN 150 (Sb (III) IC50 = 20.9 mu M) and its variation when treated with 120 mu M of Sb(III) potassium tartrate for 12 h, as well as with its Sb(III) resistant counterpart obtained by growth of the parasites under increasing Sb(III) in a step-wise manner up to 180 mu M. The number of metabolites compared were of 264 for BCN150 Sb(III) treated versus nontreated and of 195 for Sb(III) resistant versus susceptible parasites. After successive data filtering, differences in seven metabolites identified in databases for Leishmania pathways, showed the highest significant differences, corresponding mainly to amino acids or their metabolite surrogates. Most of them were assigned to sulfur containing amino acids and polyamine biosynthetic pathways, of special relevance considering the deterioration of the thiol-dependent redox metabolism in Leishmania by Sb(III). Given the low concentrations typical for most of these metabolites, the assay can be considered a success that should be explored for new biological questions.