A genetically encoded Förster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics.
A genetically encoded Förster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics.
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DOI：
10.1016/j.ab.2014.12.019
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发表时间：
2015-04-01
影响因子：
2.9
通讯作者：
Zamboni N
中科院分区：
文献类型：
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作者：
Peroza EA;Ewald JC;Parakkal G;Skotheim JM;Zamboni N
海藻糖-6-磷酸是细菌、酵母和植物中糖代谢、生长和渗透平衡的关键调节剂。为了直接观察细胞内海藻糖-6-磷酸的水平，我们开发了一系列特定的Förster共振能量转移（FRET）传感器，用于体内显微镜。我们展示了对大肠杆菌海藻糖途径调控的实时监测。在酿酒酵母中，我们可以证明在葡萄糖生长过程中游离海藻糖-6-磷酸的浓度在一个足以抑制己糖激酶的范围内。这些发现支持海藻糖-6-磷酸作为负反馈系统效应的假设，类似于哺乳动物细胞中葡萄糖-6-磷酸对己糖激酶的抑制和糖酵解通量的控制。
Trehalose-6-phosphate is a pivotal regulator of sugar metabolism, growth, and osmotic equilibrium in bacteria, yeasts, and plants. To directly visualize the intracellular levels of intracellular trehalose-6-phosphate, we developed a series of specific Förster resonance energy transfer (FRET) sensors for in vivo microscopy. We demonstrated real-time monitoring of regulation in the trehalose pathway of Escherichia coli. In Saccharomyces cerevisiae, we could show that the concentration of free trehalose-6-phosphate during growth on glucose is in a range sufficient for inhibition of hexokinase. These findings support the hypothesis of trehalose-6-phosphate as the effector of a negative feedback system, similar to the inhibition of hexokinase by glucose-6-phosphate in mammalian cells and controlling glycolytic flux.