title: Construction of a panel of glucose indicator proteins for continuous glucose monitoring.
abstract: 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 μM) within living cells through FRET imaging microscopy measurement.
authors.affiliation: Biomedical Engineering Program, College of Engineering, University of Arkansas, 203 Engineering Hall, Fayetteville, AR 72701, USA.
authors.forename: Sha
authors.initials: S
authors.lastname: Jin
authors.forename: Jithesh V
authors.initials: JV
authors.lastname: Veetil
authors.forename: Jared R
authors.initials: JR
authors.lastname: Garrett
authors.forename: Kaiming
authors.initials: K
authors.lastname: Ye
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D001786:Blood Glucose / Q000032:analysis*; D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer; D051379:Mice; D015335:Molecular Probes; D016297:Mutagenesis, Site-Directed
chemical_list: D001786:Blood Glucose; D015335:Molecular Probes