Clarification of pathway-specific inhibition by Fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies
Clarification of pathway-specific inhibition by Fourier transform ion cyclotron resonance/mass spectrometry-based metabolic phenotyping studies
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DOI：
10.1104/pp.106.080317
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发表时间：
2006-10-01
期刊：
影响因子：
7.4
通讯作者：
Ohta, Daisaku
中科院分区：
文献类型：
--
作者：
Oikawa, Akira;Nakamura, Yukiko;Ohta, Daisaku
我们开发了一种基于直接注入傅里叶变换离子回旋共振质谱 (FT-ICR/MS) 的代谢分析方案。该方案包括：（1）在优化的 FT-ICR/MS 分析条件下收集可重复的数据； (2) 使用新开发的代谢组学工具（DMASS 软件）进行自动质量误差校正和代谢组表征的多变量分析； (3) 通过搜索物种-代谢物关系数据库 KNApSAcK 鉴定候选标记代谢物； (4)通过MS/MS方法进行结构分析。该方案应用于用不同除草化学品处理的拟南芥幼苗的代谢表型，以进行途径特异性抑制。将拟南芥提取物直接注入 FT-ICR/MS 系统上的电喷雾电离源中。获得的代谢组学数据由质荷比值和经过主成分分析的离子强度信息组成，除草剂处理的代谢表型与不含除草剂处理的代谢表型明显不同。通过 KNApSAcK 数据库搜索，从每种除草剂处理中找到了代表此类代谢表型的候选代谢物。数据库搜索和 MS/MS 分析表明特定代谢物（包括几种黄酮苷）的剂量依赖性积累模式。基于 FT-ICR/MS 与 DMASS 程序相结合的代谢表型方案作为高通量代谢表型研究的通用工具进行了讨论。
We have developed a metabolic profiling scheme based on direct-infusion Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The scheme consists of: (1) reproducible data collection under optimized FT-ICR/MS analytical conditions; (2) automatic mass-error correction and multivariate analyses for metabolome characterization using a newly developed metabolomics tool (DMASS software); (3) identification of marker metabolite candidates by searching a species-metabolite relationship database, KNApSAcK; and (4) structural analyses by an MS/MS method. The scheme was applied to metabolic phenotyping of Arabidopsis (Arabidopsis thaliana) seedlings treated with different herbicidal chemical classes for pathway-specific inhibitions. Arabidopsis extracts were directly infused into an electrospray ionization source on an FT-ICR/MS system. Acquired metabolomics data were comprised of mass-to-charge ratio values with ion intensity information subjected to principal component analysis, and metabolic phenotypes from the herbicide treatments were clearly differentiated from those of the herbicide-free treatment. From each herbicide treatment, candidate metabolites representing such metabolic phenotypes were found through the KNApSAcK database search. The database search and MS/MS analyses suggested dose-dependent accumulation patterns of specific metabolites including several flavonoid glycosides. The metabolic phenotyping scheme on the basis of FT-ICR/MS coupled with the DMASS program is discussed as a general tool for high throughput metabolic phenotyping studies.