Metabolite profiling of Chlamydomonas reinhardtii under nutrient deprivation
Metabolite profiling of Chlamydomonas reinhardtii under nutrient deprivation
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DOI：
10.1104/pp.105.071589
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发表时间：
2005-12-01
期刊：
影响因子：
7.4
通讯作者：
Fiehn, O
中科院分区：
文献类型：
--
作者：
Bölling, C;Fiehn, O
开发了一种莱茵衣藻细胞代谢物分析技术，用于低分子量极性化合物的多重平行分析。实验方案经过优化，可快速灭活酶活性、实现最大提取能力并处理大量样品。通过气相色谱与飞行时间质谱联用的快速采样、提取和分析，可以测量单个样品中的 800 多种分析物，其中 100 多种分析物可以被鉴定。大多数情况下，SE 低于 10% 时即可确定分析物响应。莱茵衣藻菌株 CC-125 的野生型细胞在缺乏氮、磷、硫或铁的生长条件下会产生高度独特的代谢谱。各个代谢物的稳态水平发生显着变化。与对照条件相比，硫耗尽的细胞积累的4-羟脯氨酸超过50倍，而2-酮缬氨酸的量减少至对照水平的2%。通过主成分分析以严格的定量方式总结了每种化合物对代谢表型中观察到的差异的贡献，该方法清楚地区分了来自不同生长状态的细胞，并表明磷耗尽条件诱导的缺乏综合征与对氮、硫或铁饥饿的反应完全不同。
A metabolite profiling technique for Chlamydomonas reinhardtii cells for multiparallel analysis of low-molecular weight polar compounds was developed. The experimental protocol was optimized to quickly inactivate enzymatic activity, achieve maximum extraction capacity, and process large sample quantities. As a result of the rapid sampling, extraction, and analysis by gas chromatography coupled to time-of-flight mass spectrometry, more than 800 analytes from a single sample could be measured, of which more than 100 could be identified. Analyte responses could be determined mostly with SEs less than 10%. Wild-type cells of C. reinhardtii strain CC-125 subjected to nitrogen-, phosphorus-, sulfur-, or iron-depleted growth conditions develop highly distinctive metabolite profiles. Individual metabolites undergo marked changes in their steady-state levels. Compared to control conditions, sulfur- depleted cells accumulated 4-hydroxyproline more than 50-fold, whereas the amount of 2-ketovaline was reduced to 2% of control levels. The contribution of each compound to the differences observed in the metabolic phenotypes is summarized in a quantitatively rigorous way by principal component analysis, which clearly discriminates the cells from different growth regimes and indicates that phosphorus- depleted conditions induce a deficiency syndrome quite different from the response to nitrogen, sulfur, or iron starvation.