A genetically encoded Forster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics

作者：Peroza Estevao A; Ewald Jennifer C; Parakkal Geetha; Skotheim Jan M; Zamboni Nicola^*

来源：Analytical Biochemistry, 2015, 474: 1-7.

DOI：10.1016/j.ab.2014.12.019

• Trehalose-6-phosphate
• FRET sensor
• Glycolysis regulation

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 Forster resonance energy transfer (FRET) sensors for in vivo microscopy. We demonstrated real-time monitoring of regulation in the trehalose pathway of Escherichia con. 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.

• 出版日期2015-4-1