Helassa, Nordine ORCID: 0000-0003-3743-1886, Garnett, James P, Farrant, Matthew, Khan, Faaizah, Pickup, John C, Hahn, Klaus M, MacNevin, Christopher J, Tarran, Robert and Baines, Deborah L
(2014)
A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration.
Biochemical Journal, 464 (2).
pp. 213-220.
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Abstract
Both lung disease and elevation of blood glucose are associated with increased glucose concentration (from 0.4 to ~4.0 mM) in the airway surface liquid (ASL). This perturbation of ASL glucose makes the airway more susceptible to infection by respiratory pathogens. ASL is minute (~1 μl/cm(2)) and the measurement of glucose concentration in the small volume ASL is extremely difficult. Therefore, we sought to develop a fluorescent biosensor with sufficient sensitivity to determine glucose concentrations in ASL in situ. We coupled a range of environmentally sensitive fluorophores to mutated forms of a glucose/galactose-binding protein (GBP) including H152C and H152C/A213R and determined their equilibrium binding properties. Of these, GBP H152C/A213R-BADAN (Kd 0.86 ± 0.01 mM, Fmax/F0 3.6) was optimal for glucose sensing and in ASL increased fluorescence when basolateral glucose concentration was raised from 1 to 20 mM. Moreover, interpolation of the data showed that the glucose concentration in ASL was increased, with results similar to that using glucose oxidase analysis. The fluorescence of GBP H152C/A213R-BADAN in native ASL from human airway epithelial cultures in situ was significantly increased over time when basolateral glucose was increased from 5 to 20 mM. Overall our data indicate that this GBP is a useful tool to monitor glucose homoeostasis in the lung.
Item Type: | Article |
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Uncontrolled Keywords: | airway epithelial cell, airway surface liquid, BADAN, glucose, glucose-binding protein |
Depositing User: | Symplectic Admin |
Date Deposited: | 04 Jun 2018 15:49 |
Last Modified: | 19 Jan 2023 01:32 |
DOI: | 10.1042/BJ20141041 |
Open Access URL: | http://www.biochemj.org/content/464/2/213.long |
Related URLs: | |
URI: | https://livrepository.liverpool.ac.uk/id/eprint/3022106 |
Available Versions of this Item
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A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 25 Jul 2017 12:04)
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A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 26 Jul 2017 11:21)
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A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 02 Aug 2017 08:16)
- A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 04 Jun 2018 15:49) [Currently Displayed]
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A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 02 Aug 2017 08:16)
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A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 26 Jul 2017 11:21)