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

Helassa, N, Garnett, JP, Farrant, M, Khan, F, Pickup, JC, Hahn, KM, MacNevin, CJ, Tarran, R and Baines, DL (2014) A novel fluorescent sensor protein for detecting changes in airway surface liquid glucose concentration. Biochemical Journal, 464 (2). 213 - 220. ISSN 0264-6021, 1470-8728

Access the full-text of this item by clicking on the Open Access link.

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
Depositing User:	Symplectic Admin
Date Deposited:	25 Jul 2017 12:04
Last Modified:	19 Jan 2023 06:58
DOI:	10.1042/BJ20141041
Open Access URL:	http://www.biochemj.org/content/ppbiochemj/464/2/2...
Related URLs:	http://dx.doi.org/10.1042/BJ20141041
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3008599

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) [Currently Displayed]
  • 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)