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

, , , , , , , ,
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
Accepted
Subject Areas
Bioengineering, Biophysics, Molecular Biology
Keywords
D-glucose/D-galactose-binding protein, protein stability, fluorescent dye BADAN, biosensors, glucose binding constant
Copyright
© 2013 Fonin et al.
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

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.

Author Comment

This manuscript was submitted for review with PeerJ.

Additional Information

Competing Interests

K.K. Turoverov and I.M. Kuznetsova are Academic Editors for PeerJ.

The authors have no competing interests.

Author Contributions

Alexander V Fonin conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper.

Olga V Stepanenko performed the experiments, analyzed the data.

Olga I Povarov conceived and designed the experiments, performed the experiments, analyzed the data, wrote the paper.

Catherine A Volova performed the experiments.

Elizaveta M Filippova performed the experiments.

Grigory S Bublikov performed the experiments, analyzed the data.

Irina M. Kuznetsova conceived and designed the experiments, analyzed the data, wrote the paper.

Alexander P Demchenko conceived and designed the experiments, analyzed the data, wrote the paper.

Konstantin K Turoverov conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper.

Grant Disclosures

The following grant information was disclosed by the authors:

This work was supported in part by the Ministry of Education and Science of the Russian Federation (Agreements number 8830 and 14.132.21.311), the Scholarships from the President of RF (SP-2390.2012.4, SP-563.2012.4), the Russian Foundation for Basic Research (grant 12-04-31708 mol_a ), Program MCB RAS.

Funding

This work was supported in part by the Ministry of Education and Science of the Russian Federation (Agreements number 8830 and 14.132.21.311), the Scholarships from the President of RF (SP-2390.2012.4, SP-563.2012.4), the Russian Foundation for Basic Research (grant 12-04-31708 mol_a ), Program MCB RAS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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