Abstract |
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.
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Authors | Faaizah Khan, Tania E Saxl, John C Pickup |
Journal | Analytical biochemistry
(Anal Biochem)
Vol. 399
Issue 1
Pg. 39-43
(Apr 01 2010)
ISSN: 1096-0309 [Electronic] United States |
PMID | 19961827
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright 2009 Elsevier Inc. All rights reserved. |
Chemical References |
- 6-bromoacetyl-2-dimethylaminonaphthalene
- Blood Glucose
- Calcium-Binding Proteins
- Fluorescent Dyes
- Monosaccharide Transport Proteins
- Periplasmic Binding Proteins
- Protein Isoforms
- galactose-binding protein
- 2-Naphthylamine
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Topics |
- 2-Naphthylamine
(analogs & derivatives, chemistry)
- Amino Acid Substitution
- Biosensing Techniques
- Blood Glucose
(analysis)
- Calcium-Binding Proteins
(genetics, metabolism)
- Diabetes Mellitus
(diagnosis)
- Enzyme Assays
(methods)
- Fluorescent Dyes
(chemistry)
- Humans
- Monosaccharide Transport Proteins
(genetics, metabolism)
- Mutagenesis, Site-Directed
- Periplasmic Binding Proteins
(genetics, metabolism)
- Protein Isoforms
(genetics, metabolism)
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