Spectral properties of BADAN in solutions with different polarities

Fonin, Alexander V.; Kuznetsova, Irina M.; Turoverov, Konstantin K.

doi:10.1016/j.molstruc.2015.01.038

Cited by 4 publications

(4 citation statements)

References 44 publications

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“…The position and shape of the BADAN fluorescence excitation spectrum do not match those of the absorption spectrum of the dye and significantly depend on the fluorescence recording wavelength (Figure 2). These indicate the heterogeneity of the ensemble of BADAN molecules in the ground state [56,57]. The shape of the excitation anisotropy fluorescence spectrum of BADAN in the long-wavelength absorption band (Figure 2) indicates the collinearity of the absorption and emission dipole moment, which confirms the known data on the absence of reorientation of the dipole moment of the dye molecule upon transition to the excited state [21].…”

Section: Ggbp/h152c Apo-and Holoformssupporting

confidence: 81%

Photophysical Properties of BADAN Revealed in the Study of GGBP Structural Transitions

Fonin

Silonov

Antifeeva

et al. 2021

IJMS

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The fluorescent dye BADAN (6-bromoacetyl-2-dimetylaminonaphtalene) is widely used in various fields of life sciences, however, the photophysical properties of BADAN are not fully understood. The study of the spectral properties of BADAN attached to a number of mutant forms of GGBP, as well as changes in its spectral characteristics during structural changes in proteins, allowed to shed light on the photophysical properties of BADAN. It was shown that spectral properties of BADAN are determined by at least one non-fluorescent and two fluorescent isomers with overlapping absorbing bands. It was found that BADAN fluorescence is determined by the unsolvated “PICT” (planar intramolecular charge transfer state) and solvated “TICT” (twisted intramolecular charge transfer state) excited states. While “TICT” state can be formed both as a result of the “PICT” state solvation and as a result of light absorption by the solvated ground state of the dye. BADAN fluorescence linked to GGBP/H152C apoform is quenched by Trp 183, but this effect is inhibited by glucose intercalation. New details of the changes in the spectral characteristics of BADAN during the unfolding of the protein apo and holoforms have been obtained.

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Section: Ggbp/h152c Apo-and Holoformssupporting

confidence: 81%

Photophysical Properties of BADAN Revealed in the Study of GGBP Structural Transitions

Fonin

Silonov

Antifeeva

et al. 2021

IJMS

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“…Apparently, the recorded changes in the fluorescence characteristics of BADAN linked to GGBP/H152C in the presence of these concentrations of GdnHCl are due to the significant changes in the solvation of the dye [ 79 ]. The absence of noticeable changes in the intensity and the position of the BADAN fluorescence spectrum linked to the holo-form in this range of the GdnHCl concentrations can be explained by the fact that, in this case, BADAN is already in a sufficiently rigid environment and is largely protected from the solvent.…”

Section: Resultsmentioning

confidence: 99%

Osmolyte-Like Stabilizing Effects of Low GdnHCl Concentrations on d-Glucose/d-Galactose-Binding Protein

Fonin

Golikova

Зверева

et al. 2017

IJMS

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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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“…another fluorescent probe, BADAN (6-bromo-acetyl-2-dimethylamino-naphtalene), as the signal transducing component of the biosensor. Here, the E166C-based mutants were conjugated with a BADAN to give the E166Cb and E166Cb/N170Q respectively (S1 Fig) . BADAN is a fluorescent solvatochromic dye [26]. Our rationale in exploiting BADAN is based on its high sensitivity to the microenvironment change.…”

Section: Plos Onementioning

confidence: 99%

“…Here, the E166C-based mutants were conjugated with a BADAN to give the E166Cb and E166Cb/N170Q respectively ( S1 Fig ). BADAN is a fluorescent solvatochromic dye [ 26 ]. Our rationale in exploiting BADAN is based on its high sensitivity to the microenvironment change.…”

Section: Introductionmentioning

confidence: 99%

BADAN-conjugated β-lactamases as biosensors for β-lactam antibiotic detection

Au¹,

Tsang²,

Chen³

et al. 2020

PLoS ONE

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β-Lactam antibiotic detection has significant implications in food safety control, environmental monitoring and pharmacokinetics study. Here, we report the development of two BADAN-conjugated β-lactamases, E166Cb and E166Cb/N170Q, as sensitive biosensors for β-lactam antibiotic detection. These biosensors were constructed by coupling an environment-sensitive BADAN probe onto location 166 at the active site of the PenP β-lactamase E166C and E166C/N170Q mutants. They gave fluorescence turn-on signals in response to β-lactam antibiotics. Molecular dynamics simulation of E166Cb suggested that the turn-on signal might be attributed to a polarity change of the microenvironment of BADAN and the removal of the fluorescence quenching effect on BADAN exerted by a nearby Tyr-105 upon the antibiotic binding. In the detection of four β-lactams (penicillin G, penicillin V, cefotaxime and moxalactam), both E166Cb and E166Cb/N170Q delivered signal outputs in an antibiotic-concentration dependent manner with a dynamic range spanning from 10 nM to 1 μM. Compared to E166Cb, E166Cb/N170Q generally exhibited more stable signals owing to its higher deficiency in hydrolyzing the antibiotic analyte. The overall biosensor performance of E166Cb and E166Cb/N170Q was comparable to that of their respective fluorescein-modified counterparts, E166Cf and E166Cf/N170Q. But comparatively, the BADAN-conjugated enzymes showed a higher sensitivity, displayed a faster response in detecting moxalactam and a more stable fluorescence signals towards penicillin G. This study illustrates the potential of BADAN-conjugated β-lactamases as biosensing devices for β-lactam antibiotics.

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Spectral properties of BADAN in solutions with different polarities

Cited by 4 publications

References 44 publications

Photophysical Properties of BADAN Revealed in the Study of GGBP Structural Transitions

Photophysical Properties of BADAN Revealed in the Study of GGBP Structural Transitions

Osmolyte-Like Stabilizing Effects of Low GdnHCl Concentrations on d-Glucose/d-Galactose-Binding Protein

BADAN-conjugated β-lactamases as biosensors for β-lactam antibiotic detection

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