A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration

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
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:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3022106

Available Versions of this Item

• A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 25 Jul 2017 12:04)
  • A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. (deposited 26 Jul 2017 11:21)
    • 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]