GC-EI-TOF-MS analysis of in vivo carbon-partitioning into soluble metabolite pools of higher plants by monitoring isotope dilution after 13CO2 labelling
GC-EI-TOF-MS analysis of in vivo carbon-partitioning into soluble metabolite pools of higher plants by monitoring isotope dilution after 13CO2 labelling
复制标题
DOI：
10.1016/j.phytochem.2007.03.026
复制
发表时间：
2007-08-01
期刊：
影响因子：
3.8
通讯作者：
Kopka, Joachim
中科院分区：
文献类型：
--
作者：
Huege, Jan;Sulpice, Ronan;Kopka, Joachim
建立了植物组织可溶性极性代谢物的GC-EI-TOF-MS分析方法，并应用于高等植物的动力学代谢表型分析。三甲基甲硅烷基化和甲氧基胺化代谢物衍生物的约100个典型GC-EI-MS质量片段在结构上进行了解释，用于质量同位素异构体分析，从而使已鉴定代谢物的动力学研究以及尚未鉴定代谢物的所谓官能团监测成为可能。使用用C-13最大标记的拟南芥Col-0或水稻IR 57111植物优化（CO2）-C-13标记后同位素稀释的监测。碳同位素稀释进行了评估，短期（2小时）和长期（3天）的动力学测量代谢物池在根和芽。这两种方法都被证明能够表征代谢物特异性分配过程和动力学。采用简化数据简化方案，包括从质量同位素分布计算C-13富集和初始C-13稀释率。二甲双胍在其各自的代谢产物中表现出高度多样化的代谢产物和C-13标记的器官特异性半衰期（C-13半衰期）。这一观察结果意味着为最佳动力学监测设定代谢物特定时间段。本文提出了一种高等植物代谢动力学表型分析的通用实验设计。(C)2007爱思唯尔有限公司保留所有权利。
The established GC-EI-TOF-MS method for the profiling of soluble polar metabolites from plant tissue was employed for the kinetic metabolic phenotyping of higher plants. Approximately 100 typical GC-EI-MS mass fragments of trimethylsilylated and methoxyaminated metabolite derivatives were structurally interpreted for mass isotopomer analysis, thus enabling the kinetic study of identified metabolites as well as the so-called functional group monitoring of yet non-identified metabolites. The monitoring of isotope dilution after (CO2)-C-13 labelling was optimized using Arabidopsis thaliana Col-0 or Oryza sativa IR57111 plants, which were maximally labelled with C-13. Carbon isotope dilution was evaluated for short (2 h) and long-term (3 days) kinetic measurements of metabolite pools in root and shoots. Both approaches were shown to enable the characterization of metabolite specific partitioning processes and kinetics. Simplifying data reduction schemes comprising calculation of C-13-enrichment from mass isotopomer distributions and of initial C-13-dilution rates were employed. Metabolites exhibited a highly diverse range of metabolite and organ specific half-life of C-13-label in their respective Pools (C-13-half-life). This observation implied the setting of metabolite specific periods for optimal kinetic monitoring. A current experimental design for the kinetic metabolic phenotyping of higher plants is proposed. (C) 2007 Elsevier Ltd. All rights reserved.