Comparison of quantitative metabolite imaging tools and carbon-13 techniques for fluxomics.
Comparison of quantitative metabolite imaging tools and carbon-13 techniques for fluxomics.
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DOI：
10.1007/978-1-60327-563-7_19
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发表时间：
2009
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影响因子：
--
通讯作者：
Frommer, Wolf B
中科院分区：
文献类型：
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作者：
Niittylae, Totte;Chaudhuri, Bhavna;Sauer, Uwe;Frommer, Wolf B
分析技术的最新发展允许真实的实时快速分析代谢。通量组学旨在确定通过代谢或信号通路参与通量调节的基因。通过代谢或信号传导途径的通量由其单个组分的活性决定;调节可以在许多水平发生，包括转录、翻译后和变构水平。目前有两种技术用于监测通量。第一种是用示踪剂如C13对生物体进行脉冲标记，然后对标记物分配到不同化合物中进行质谱分析。第二种方法是基于使用通量传感器，蛋白质的构象变化，以配体结合。荧光共振能量转移（FRET）检测构象变化，并作为配体浓度的代理。这两种方法都具有很高的时间分辨率。与质谱分析相反，FRET纳米传感器仅监测单一化合物，但FRET纳米传感器的优点是它们产生具有细胞和亚细胞分辨率的数据。
The recent development of analytic technologies allows fast analysis of metabolism in real time. Fluxomics aims to define the genes involved in regulation of flux through a metabolic or signaling pathway. Flux through a metabolic or signaling pathway is determined by the activity of its individual components; regulation can occur at many levels, including transcriptional, posttranslational, and allosteric levels. Currently two technologies are used to monitor fluxes. The first is pulse labeling of the organism with a tracer such as C13, followed by mass spectrometric analysis of the partitioning of label into different compounds. The second approach is based on the use of flux sensors, proteins that respond with a conformational change to ligand binding. Fluorescence resonance energy transfer (FRET) detects the conformational change and serves as a proxy for ligand concentration. Both methods provide high time resolution. In contrast to mass spectrometry assays, FRET nanosensors monitor only a single compound, but the advantage of FRET nanosensors is that they yield data with cellular and subcellular resolution.