Diabetes Research Group, King's College London School of Medicine, Guy's Hospital, London SE1 1UL, UK.
Anal Biochem. 2010 Apr 1;399(1):39-43. doi: 10.1016/j.ab.2009.11.035. Epub 2009 Dec 2.
We synthesized mutants of glucose/galactose-binding protein (GBP), labeled with the environmentally sensitive fluorophore Badan, with the aim of producing a fluorescence-based glucose sensing system with an operating range compatible with continuous glucose monitoring in patients with diabetes mellitus. From five mutants tested, the triple mutant H152C/A213R/L238S-Badan showed a large (200%) maximal increase in fluorescence intensity on the addition of glucose, with a binding constant (K(d)) of 11 mM, an operating range of approximately 1-100 mM, and similar responses in buffer and serum. The mean fluorescence lifetime of this mutant also increased by 70% on the addition of glucose. We conclude that the GBP mutant H152C/A213R/L238S, when labeled with Badan, is suitable for development as a robust sensor for in vivo glucose monitoring in diabetes.
我们合成了葡萄糖/半乳糖结合蛋白 (GBP) 的突变体，用环境敏感荧光染料 Badan 进行标记，目的是开发一种基于荧光的葡萄糖传感系统，其工作范围与糖尿病患者的连续血糖监测相匹配。在测试的五个突变体中，三重突变体 H152C/A213R/L238S-Badan 在加入葡萄糖时表现出荧光强度的大幅增加（200%），结合常数（K(d)）为 11mM，工作范围约为 1-100mM，在缓冲液和血清中的反应相似。该突变体的平均荧光寿命在加入葡萄糖后也增加了 70%。我们得出结论，当用 Badan 标记时，GBP 突变体 H152C/A213R/L238S 适合开发为用于糖尿病体内葡萄糖监测的稳健传感器。
