Hu Hanyang, Wei Yufeng, Wang Daocheng, Su Ni, Chen Xianjun, Zhao Yuzheng, Liu Guixia, Yang Yi
Synthetic Biology and Biotechnology Laboratory, State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing Technology, East China University of Science and Technology 130 Mei Long Road Shanghai 200237 China.
Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology 130 Mei Long Road Shanghai 200237 China
RSC Adv. 2018 Jan 10;8(5):2485-2489. doi: 10.1039/c7ra11347a. eCollection 2018 Jan 9.
Glucose is the main source of energy and carbon in organisms and plays a central role in metabolism and cellular homeostasis. However, the sensitive fluctuation of glucose in living cells is difficult to monitor. Thus, we developed a series of ratiometric, highly responsive, single fluorescent protein-based glucose sensors of wide dynamic range by combining a circularly permuted yellow fluorescent protein with a bacterial periplasmic glucose/galactose-binding protein. We used these sensors to monitor glucose transport in living cells, and found that the cells take up glucose within 10 min to maintain physiological glucose levels, and observed the differences in glucose uptake and glucose metabolism between wild-type and Mlc knockout cells. These sensors can be specific and simple tools for glucose detection and non-invasive tools for real-time monitoring of glucose metabolism .
葡萄糖是生物体中能量和碳的主要来源,在新陈代谢和细胞内稳态中起着核心作用。然而,活细胞中葡萄糖的敏感波动难以监测。因此,我们通过将环状排列的黄色荧光蛋白与细菌周质葡萄糖/半乳糖结合蛋白相结合,开发了一系列基于单一荧光蛋白的比率型、高响应性、宽动态范围的葡萄糖传感器。我们使用这些传感器监测活细胞中的葡萄糖转运,发现细胞在10分钟内摄取葡萄糖以维持生理葡萄糖水平,并观察到野生型和Mlc基因敲除细胞之间葡萄糖摄取和葡萄糖代谢的差异。这些传感器可以成为用于葡萄糖检测的特异性和简单工具,以及用于实时监测葡萄糖代谢的非侵入性工具。