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

Alexander V Fonin; Olga V Stepanenko; Olga I Povarov; Catherine A Volova; Elizaveta M Filippova; Grigory S Bublikov; Irina M. Kuznetsova; Alexander P Demchenko; Konstantin K Turoverov

doi:10.7287/peerj.preprints.111v1

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

Alexander V Fonin¹, Olga V Stepanenko¹, Olga I Povarov¹, Catherine A Volova^1,2, Elizaveta M Filippova^1,2, Grigory S Bublikov¹, Irina M. Kuznetsova^1,3, Alexander P Demchenko⁴, Konstantin K Turoverov ¹

1 Laboratory of Structural Dynamics, Stability and Folding of Proteins, Institute of Cytology, Russian Academy of Science, St.Petersburg, Russia

2 Department of Physical Electronics, St. Petersburg State Polytechnical University, St.Petersburg, Russia

3 Department of Biophysics, St. Petersburg State Polytechnical University, St.Petersburg, Russia

4 Laboratory of Nanobiotechnologies, Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine, Kiev, Ukraine

DOI: 10.7287/peerj.preprints.111v1

Published: 2013-11-22
Accepted: 2013-11-22

Subject Areas: Bioengineering, Biophysics, Molecular Biology
Keywords: D-glucose/D-galactose-binding protein, protein stability, fluorescent dye BADAN, biosensors, glucose binding constant

Licence: This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Cite this article: Fonin AV, Stepanenko OV, Povarov OI, Volova CA, Filippova EM, Bublikov GS, Kuznetsova IM, Demchenko AP, Turoverov KK. 2013. Spectral characteristics of the mutant protein D-glucose/D-galactose-binding protein GGBP/H152C with an attached fluorescent dye BADAN: Influence of external conditions. PeerJ PrePrints 1:e111v1 https://doi.org/10.7287/peerj.preprints.111v1

Abstract

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.

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