Spectral characteristics of the mutant protein D-glucose/D-galactose-binding protein GGBP/H152C with an attached fluorescent dye BADAN: Influence of external conditions

The mutant form of the D-glucose/D-galactose-binding protein GGBP/H152C with the solvatochromic dye BADAN attached to cysteine residue Cys 152 can be used as a potential base for a sensitive element of glucose biosensor system. We investigated the influence of various factors on the physical-chemica...

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Bibliographic Details
Published in:PeerJ preprints 2013-11
Main Authors: Fonin, Alexander V, Stepanenko, Olga V, Povarov, Olga I, Volova, Catherine A, Filippova, Elizaveta M, Bublikov, Grigory S, Kuznetsova, Irina M, Demchenko, Alexander P, Turoverov, Konstantin K
Format: Article
Language:English
Subjects:
Biosensors
D-Galactose
Denaturation
Dyes
Fluorescent indicators
Galactose
Glucose
Ionic strength
pH effects
Protein D
Proteins
Sugar
Viscosity
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Summary:The mutant form of the D-glucose/D-galactose-binding protein GGBP/H152C with the solvatochromic dye BADAN attached to cysteine residue Cys 152 can be used as a potential base for a sensitive element of glucose biosensor system. We investigated the influence of various factors on the physical-chemical properties of GGBP/H152C-BADAN and on its complexation with glicose . The high affinity (K d = 8.5 μM) and rapid binding of glucose ( even in solutions with a viscosity value of 4 cP, the formation time of the protein-glucose is not longer than three seconds ) allows for theuse of GGBP/H152C-BADAN to determine the sugar content in biological fluids extracted using transdermal techniques. It was shown that the changes in the ionic strength and pH within the physiological range did not have a significant influence on the fluorescent characteristics of GGBP/H152C-BADAN. The mutant form GGBP/H152C has relatively low resistance to denaturation action. This result emphasizes the need to find more stable proteins for the creation of a sensitive element for a glucose biosensor system.
ISSN:2167-9843
DOI:10.7287/peerj.preprints.111v1