D-glucose/d-galactose-binding Protein, Guanidine Hydrochloride, Osmolyte-like Stabilizing Effect, Fluorescent Label BADAN, Protein Conformers, Protein Function, Conformational Ensemble
Digital Object Identifier (DOI)
The ability of d-glucose/d-galactose-binding protein (GGBP) to reversibly interact with its ligands, glucose and galactose, makes this protein an attractive candidate for sensing elements of glucose biosensors. This potential is largely responsible for attracting researchers to study the conformational properties of this protein. Previously, we showed that an increase in the fluorescence intensity of the fluorescent dye 6-bromoacetyl-2-dimetylaminonaphtalene (BADAN) is linked to the holo-form of the GGBP/H152C mutant in solutions containing sub-denaturing concentrations of guanidine hydrochloride (GdnHCl). It was hypothesized that low GdnHCl concentrations might lead to compaction of the protein, thereby facilitating ligand binding. In this work, we utilize BADAN fluorescence spectroscopy, intrinsic protein UV fluorescence spectroscopy, and isothermal titration calorimetry (ITC) to show that the sub-denaturing GdnHCl concentrations possess osmolyte-like stabilizing effects on the structural dynamics, conformational stability, and functional activity of GGBP/H152C and the wild type of this protein (wtGGBP). Our data are consistent with the model where low GdnHCl concentrations promote a shift in the dynamic distribution of the protein molecules toward a conformational ensemble enriched in molecules with a tighter structure and a more closed conformation. This promotes the increase in the configurational complementarity between the protein and glucose molecules that leads to the increase in glucose affinity in both GGBP/H152C and wtGGBP.
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Citation / Publisher Attribution
International Journal of Molecular Sciences, v. 18, issue 9, art. 2008
Scholar Commons Citation
Fonin, Alexander V.; Golikova, Alexandra D.; Zvereva, Irina A.; D’Auria, Sabato; Staiano, Maria; Uversky, Vladimir N.; Kuznetsova, Irina M.; and Turoverov, Konstantin K., "Osmolyte-Like Stabilizing Effects of Low GdnHCl Concentrations on d-Glucose/d-Galactose-Binding Protein" (2017). Molecular Medicine Faculty Publications. 848.