Spectral characteristics of the mutant form GGBP/H152C of D-glucose/D-galactose-binding protein labeled with fluorescent dye BADAN: influence of external factors.

Abstract:

:The mutant form GGBP/H152C of the D-glucose/D-galactose-binding protein with the solvatochromic dye BADAN linked 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 external factors on the physical-chemical properties of GGBP/H152C-BADAN and its complex with glucose. The high affinity (Kd = 8.5 µM) and high binding rate of glucose make GGBP/H152C-BADAN a good candidate to determine the sugar content in biological fluids extracted using transdermal techniques. It was shown that changes in the ionic strength and pH of solution 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 of GdnHCl and urea. This result emphasizes the need to find more stable proteins for the creation of a sensitive element for a glucose biosensor system.

journal_name

PeerJ

journal_title

PeerJ

authors

Fonin AV,Stepanenko OV,Povarova OI,Volova CA,Philippova EM,Bublikov GS,Kuznetsova IM,Demchenko AP,Turoverov KK

doi

10.7717/peerj.275

subject

Has Abstract

pub_date

2014-03-18 00:00:00

pages

e275

issn

2167-8359

pii

275

journal_volume

2

pub_type

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