Russian Research Center Kurchatov Institute, pl. Akademika Kurchatova 1, 123182 Moscow, Russia.
Biochemistry (Mosc). 2012 Aug;77(8):864-77. doi: 10.1134/S000629791208007X.
In contrast to the abundance of systems-oriented approaches describing changes on the transcriptome or proteome level, relatively few studies have employed the metabolome. The goal of the presented research was to identify as many intracellular metabolites as possible in a Spiroplasma melliferum extract by flow injection time-of-flight mass spectrometry. The Mollicutes class bacterium S. melliferum is a member of a unique category of bacteria that have in common the absence of a cell wall, a reduced genome, and simplified metabolic pathways. Metabolite identification was confirmed by fragmentation of previously detected ions by target mass spectrometry. The selected liquid chromatography approach, hydrophilic interaction chromatography with amino and silica columns, effectively separates highly polar cellular metabolites prior to their detection on a high accuracy mass spectrometer in positive and negative acquisition mode for each column. Here we present reliable measurement of 76 metabolites, including components of sugar, amino acid, and nucleotide metabolism. We have identified about a third of the possible intracellular S. melliferum metabolites predicted by genome annotation.
与大量描述转录组或蛋白质组水平变化的系统方法相比，相对较少的研究采用了代谢组学。本研究的目的是通过流动注射飞行时间质谱法尽可能多地鉴定出蜜环菌提取物中的细胞内代谢物。柔膜体纲细菌蜜环菌是一种独特的细菌，其共同特征是缺乏细胞壁、基因组较小和代谢途径简化。通过对先前检测到的离子进行靶向质谱碎裂，确认了代谢物的鉴定。所选的液相色谱方法，即带有氨基和硅胶柱的亲水作用色谱法，在正、负离子采集模式下，在高精度质谱仪上检测之前，可有效分离高度极性的细胞代谢物。在此，我们提出了可靠的测量方法，可测量 76 种代谢物，包括糖、氨基酸和核苷酸代谢的成分。我们已经鉴定出了约三分之一的通过基因组注释预测的可能的蜜环菌细胞内代谢物。
