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基于绿色荧光蛋白的葡萄糖指示剂可报告活细胞中的葡萄糖动态。

Green Fluorescent Protein-Based Glucose Indicators Report Glucose Dynamics in Living Cells.

机构信息

Department of Life Sciences, Graduate School of Arts and Sciences , The University of Tokyo , 3-8-1 Komaba , Meguro , Tokyo 153-8902 , Japan.

Department of Biological Sciences, Graduate School of Science , The University of Tokyo , 7-3-1 Hongo , Bunkyo , Tokyo 113-0033 , Japan.

出版信息

Anal Chem. 2019 Apr 2;91(7):4821-4830. doi: 10.1021/acs.analchem.9b00447. Epub 2019 Mar 22.

Abstract

Glucose is the most important energy source for living animals. Here, we developed a series of single fluorescent protein (FP)-based glucose indicators, named as "Green Glifons", to understand the hierarchal and mutual relationships between molecules involved in energy metabolism. Three indicators showed a different EC for glucose (50, 600, and 4000 μM), producing a ∼7-fold change in fluorescence intensity in response to glucose. The indicators could visualize glucose dynamics in the cytoplasm, plasma membrane, nucleus and mitochondria of living HeLa cells and in vivo, in the pharyngeal muscle of C. elegans and could measure murine blood glucose levels. Finally, the indicators were applicable to dual-color imaging, revealing the dynamic interplay between glucose and Ca in mouse pancreatic MIN6 m9 β cells. We propose that these indicators will facilitate and contribute to in vivo and multicolor imaging of energy metabolism.

摘要

葡萄糖是活体动物最重要的能量来源。在这里,我们开发了一系列基于单荧光蛋白 (FP) 的葡萄糖指示剂,命名为“Green Glifons”,以了解参与能量代谢的分子之间的层次和相互关系。三种指示剂对葡萄糖的 EC 不同(50、600 和 4000 μM),对葡萄糖的荧光强度有 7 倍的变化。这些指示剂可以可视化活 HeLa 细胞细胞质、质膜、核和线粒体以及活体秀丽隐杆线虫咽肌中的葡萄糖动态,并可测量鼠血葡萄糖水平。最后,这些指示剂可用于双色成像,揭示了小鼠胰腺 MIN6 m9β细胞中葡萄糖与 Ca 之间的动态相互作用。我们提出,这些指示剂将有助于并促进能量代谢的体内和多色成像。

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