Development of red genetically encoded biosensor for visualization of intracellular glucose dynamics
Development of red genetically encoded biosensor for visualization of intracellular glucose dynamics
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DOI：
10.1016/j.chembiol.2021.06.002
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发表时间：
2022-01-20
影响因子：
8.6
通讯作者：
Tsuboi, Takashi
中科院分区：
文献类型：
--
作者：
Mita, Marie;Sugawara, Izumi;Tsuboi, Takashi
葡萄糖是生物体的主要能量来源，了解细胞内葡萄糖的时空动力学具有重要意义。一种基于单一荧光蛋白的葡萄糖指示剂，名为“Red Glifons”，已经被开发出来，适用于活细胞和双色成像。在10 mM葡萄糖存在下，这些指示剂的荧光强度增加了3倍以上。开发的两种Red Glifons对葡萄糖具有不同的半最大有效浓度(EC50)值(300µM和3000µM)，并能够监测广泛的葡萄糖动态。红色Glifon与绿色指示器相结合，可直观显示葡萄糖与ATP、乳酸或丙酮酸之间的相互作用。使用Red Glifons观察到秀丽线虫咽部肌肉、肠内分泌细胞和人iPS细胞来源的心肌细胞的葡萄糖内流。因此，这些红色葡萄糖指示剂可作为研究能量代谢中复杂相互作用的多色成像工具包。
Glucose is the main source of energy for organisms, and it is important to understand the spatiotemporal dynamics of intracellular glucose. Single fluorescent protein-based glucose indicators, named "Red Glifons'' have been developed that apply to live-cell and dual-color imaging. These indicators exhibited more than 3-fold increase in fluorescence intensity in the presence of 10 mM glucose. The two Red Glifons developed have different half-maximal effective concentration (EC50) values for glucose (300 mu M and 3,000 mu M) and are able to monitor a wide range of glucose dynamics. Red Glifon combined with green indicators allowing visualization of the interplay between glucose and ATP, lactate, or pyruvate. Glucose influx in the pharyngeal muscle of Caenorhabditis elegans, enteroendocrine cells, and human iPS cell-derived cardiac myocytes was observed using the Red Glifons. Thus these red glucose indicators serve as a multi-color imaging toolkit for investigating complex interactions in energy metabolism.