期刊
BIOSENSORS & BIOELECTRONICS
卷 26, 期 8, 页码 3427-3431出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2011.01.017
关键词
Glucose sensor; Glucose binding protein; Glucose indicator protein; FRET imaging microscopy measurement; Continuous glucose monitoring; Site-directed mutagenesis of GBP
类别
资金
- NIH [EB006378-01]
- Arkansas Biosciences Institute [0402-27504-21-726]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [0756455] Funding Source: National Science Foundation
The development of implantable glucose sensors for use in diabetes treatment has been pursued for decades. However, enzyme-based glucose sensors often fail in vivo. In our previous work, we engineered a novel glucose indicator protein (GIP) that can sense glucose without relying on any enzymes and cofactors. Nevertheless, this GIP is unsuitable for blood glucose monitoring due to its low dissociation constant. Here, we report a novel approach to creating a new GIP that can be used to monitor blood glucose level. By disrupting pi-pi stacking around GIP's glucose binding site through site-directed mutagenesis, we showed that GIP's dissociation constant can be manipulated from 0.026 mM to 7.86 mM. This approach yielded four GIP mutants. We showed that one of the mutants can be used to detect glucose from 0 to 32 mM, while another mutant can be employed to visualize intracellular glucose (0-200 mu M) within living cells through FRET imaging microscopy measurement. (C) 2011 Elsevier B.V. All rights reserved.
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