Spectroscopic studies on the interaction between disperse blue SBL and bovine serum albumin

The interaction of disperse blue SBL (DBSBL) with bovine serum albumin (BSA) was investigated using fluorescence, UV-visible and far-UV circular dichroism (CD) spectroscopy. The results showed that the fluorescence of BSA was quenched by DBSBL through static quenching after correcting for the inner filter effects (IFE). The binding constant K_b of DBSBL with BSA at 288, 298 and 303 K were 0.116×10⁶, 3.18×10⁶ and 12.3×10⁶ L mol⁻¹, respectively. The thermodynamic parameters, standard enthalpy change (ΔH⁰) and standard entropy change (ΔS⁰), for the reaction were evaluated to be 227.2 kJ mol⁻¹ and 886 J mol⁻¹ K⁻¹ according to the van’t Hoff equation. The above data suggested that the forces acting between DBSBL and BSA were predominantly hydrophobic interactions. The results of UV-visible absorption and far-UV CD spectroscopy also revealed that the conformation and microenvironment of BSA molecule were changed after DBSBL binding to BSA. At 288 K one binding site was present but at higher temperatures a second binding site was detected between DBSBL and the BSA molecule. The lower bound for the distance between the bound dye and the Trp residue is 2.35 nm as calculated from Forster energy transfer.     

Spectroscopic studies on the binding of barbital to bovine serum albumin

In this paper, the interaction between barbital and bovine serum albumin (BSA) was investigated by the method of fluorescence spectroscopy under simulative physiological conditions. Fluorescence data revealed that the fluorescence quenching of BSA by barbital was the result of the formation of BSA-barbital complex, and the effective quenching constants (K_a) were 1.468×10⁴, 1.445×10⁴ and 1.403×10⁴ M⁻¹ at 297, 303 and 310 K, respectively. The thermodynamic parameters enthalpy change (ΔH) and entropy change (ΔS) for the reaction were calculated to be −2.679 kJ mol⁻¹ and 70.76 J mol⁻¹ K⁻¹, respectively, according to the van’t Hoff equation. The results indicated that hydrophobic and electrostatic interactions were the dominant intermolecular force in stabilizing the complex. The results of synchronous fluorescence spectra showed that binding of barbital with BSA can induce conformational changes in BSA. In addition, the effects of Cu²⁺ and Zn²⁺ on the constants of BSA-barbital complex were also discussed.     

Spectrofluorimetric study on the interaction between antimicrobial drug sulfamethazine and bovine serum albumin

The interaction between the antimicrobial drug sulfamethazine (STM) and bovine serum albumin (BSA) has been studied using steady state and synchronous fluorescence spectroscopy. Fluorescence emission data revealed that BSA (2×10⁻⁶ M) fluorescence was statically quenched by STM at various concentrations, which implies that STM-BSA complex has been formed. The fluorescence emission data was analyzed via applying the Stern-Volmer analysis in combination with thermodynamic investigation, where obtained results revealed that quenching is static with quenching constants of 2.371, 1.658, and 0.916×10⁵ M⁻¹ at 298, 304, and 310 K, respectively. Binding constants and number of binding sites at different temperatures were also determined by applying the Scatchard method, which in turn were used to construct the van't Hoff plot in order to estimate the enthalpy (ΔH) and entropy changes (ΔS) for the complexation process. An average of 1.00±0.17 was estimated for the number of sites of BSA, which indicated that STM binds to BSA with stoichiometric ratio of 1:1. The values that were estimated from the van't Hoff plot for ΔH and (ΔS) were −36.8 kJ mol⁻¹ and −14.9 J mol⁻¹ K⁻¹, respectively, which indicate that the STM-BSA complex is stabilized with hydrogen bonds and van der Waals interactions. Synchronous fluorescence data was obtained at Δλ of 15 and 60 nm, where obtained results confirmed that STM binds to BSA at the tryptophan residue (Trp. 213). In addition, the distance between STM and the Trp. 213 was estimated via employing the Förster's non-radiative energy-transfer theory, and was found to be 2.73 nm, which in turn indicated that STM can bind to BSA with high probability.     

Spectroscopic and voltammetric characterizations of the interaction of two local anesthetics with bovine serum albumin

The interactions of bovine serum albumin (BSA) with two local anesthetics, procaine hydrochloride (PCH) and tetracaine hydrochloride (TCH) were studied using spectroscopic methods such as fluorescence and ultraviolet visible (UV-vis), and electrochemical techniques including cyclic voltammetry (CV) and differential pulsed stripping voltammetry (DPSV). The results obtained from these techniques turned out that both PCH and TCH could bind to BSA. The binding constants (K_A) and the number of binding sites (n) of the two drugs with BSA at different temperatures were determined, respectively. At 291 K, K_A was found as 2.40×10⁴ and 1.42×10⁴ L mol⁻¹ and n was 1.03 and 0.99 for PCH-BSA and TCH-BSA, respectively. According to van’t Hoff equation, the thermodynamic parameters, ΔG, ΔH and ΔS, were obtained, showing the involvement of hydrophobic and electrostatic force in these interactions. Based on the theory of the Förster energy transference, the distance between the acceptor (PCH or TCH) and the donor (BSA) were determined as 2.32 and 3.62 nm for PCH and TCH, respectively. The effects of Fe³⁺, Cu²⁺, Mg²⁺, Mn²⁺, Zn²⁺ and Ca²⁺ on the binding of PCH or TCH to BSA were also evaluated.     

Study on the interaction between promethazine hydrochloride and bovine serum albumin by fluorescence spectroscopy

The interaction between promethazine hydrochloride (PMT) and bovine serum albumin (BSA) in vitro was investigated by means of fluorescence spectroscopy and absorption spectroscopy. The fluorescence of BSA was quenched remarkably by PMT and the quenching mechanism was considered as static quenching by forming a complex. The association constants K_a and the number of binding sites n were calculated at different temperatures. The BSA-PMT binding distance was determined to be less than 8 nm, suggesting that energy transfer from BSA to PMT may occur. The thermodynamic parameters of the interaction between PMT and BSA were measured according to the van’t Hoff equation. The enthalpy change (ΔH) and entropy change (ΔS) were calculated to be −23.62 kJ mol⁻¹ and −0.10 J mol⁻¹ K⁻¹, respectively, which indicated that the interaction of PMT with BSA was driven mainly by van der Waals forces and hydrogen bonds. The binding process was a spontaneous process in which Gibbs free energy change (ΔG) was negative. In addition, the results of synchronous fluorescence spectra and three-dimensional fluorescence spectra showed that binding of PMT with BSA can induce conformational changes in BSA.     

Investigation of the interaction between trazodone hydrochloride and bovine serum albumin

In this paper, the binding of trazodone hydrochloride (TZH) to bovine serum albumin (BSA) was investigated by spectroscopic (fluorescence, spectrophotometry and circular dichroism) techniques under simulative physiological conditions. A strong fluorescence quenching reaction of TZH to BSA was observed and the quenching mechanism was suggested as dynamic quenching according to the Stern-Volmer equation. The binding constants of TZH with BSA at 288, 302 and 309 K were calculated as (1.56±0.003)×10⁴, (2.31±0.002)×10⁴ and (5.44±0.004)×10⁴ M⁻¹, respectively. The thermodynamic parameters, ΔH⁰ and ΔS⁰ were obtained to be 39.86±0.008 kJ mol⁻¹ and 217.89±0.011 J mol⁻¹ K⁻¹, respectively, which indicated the presence of hydrophobic forces between TZH and BSA. The spectral results observed showed that the binding of TZH to BSA induced conformational changes in BSA. Based on the Förster's theory of non-radiation energy transfer, the binding average distance, r between donor (BSA) and acceptor (TZH) was found to be 2.4 nm. The effect of common ions on binding of TZH to BSA was also examined.     

Study on the interaction of La with bovine serum albumin at molecular level

The interaction of La³⁺ to bovine serum albumin (BSA) has been investigated mainly by fluorescence spectra, UV-vis absorption spectra, and circular dichroism (CD) under simulative physiological conditions. Fluorescence data revealed that the quenching mechanism of BSA by La³⁺ was a static quenching process and the binding constant is 1.75×10⁴ L mol⁻¹ and the number of binding sites is 1 at 289 K. The thermodynamic parameters (ΔH=−20.055 kJ mol⁻¹, ΔG=−23.474 kJ mol⁻¹, and ΔS=11.831 J mol⁻¹ K⁻¹) indicate that electrostatic effect between the protein and the La³⁺ is the main binding force. In addition, UV-vis, CD, and synchronous fluorescence results showed that the addition of La³⁺ changed the conformation of BSA.     

Probing the interaction of flower-like CdSe nanostructure particles targeted to bovine serum albumin using spectroscopic techniques

The interaction between flower-like CdSe nanostructure particles (CdSe NP) and bovine serum albumin (BSA) was investigated from a spectroscopic angle under simulative physiological conditions. Under pH 7.4, CdSe NP could effectively quench the intrinsic fluorescence of BSA via static quenching. The binding constant (K_A) was 6.38, 3.27, and 1.90×10⁴ M⁻¹ at 298, 304, and 310 K, respectively and the number of binding sites was 1.20. According to the Van't Hoff equation, the thermodynamic parameters (ΔH°=−77.48 kJ mol⁻¹, ΔS°=−168.17 J mol⁻¹ K⁻¹) indicated that hydrogen bonds and van der Waals forces played a major role in stabilizing the BSA−CdSe complex. Besides, UV-vis and circular dichroism (CD) results showed that the addition of CdSe NP changed the secondary structure of BSA and led to a decrease in α-helix. These results suggested that BSA underwent substantial conformational changes induced by flower-like CdSe nanostructure particles.     

Spectroscopic studies on the interaction and sonodynamic damage of neutral red (NR) to bovine serum albumin (BSA)

In this paper, the interaction of neutral red (NR) with bovine serum albumin (BSA) and the sonodynamic damage to BSA under ultrasonic irradiation was studied by means of ultraviolet-visible (UV-vis) and fluorescence spectra. The quenching constant (K_SV=5.749×10⁴ L/mol), binding constant (K_A=3.19×10⁴ L/mol) and binding site number (n=0.9462) were measured. The binding distance (r=2.47 nm) between NR and BSA was obtained according to Föster’s non-radiative energy transfer theory. The damage process of BSA molecules was detected by the hyperchromic effect of UV-vis spectra and quenching of intrinsic fluorescence spectra. In addition, the influencing factors such as ultrasonic irradiation time and NR concentration on the damage to BSA molecules were also considered. The results showed that the damage degree is enhanced with the increase of ultrasonic irradiation time and NR concentration. The possible mechanism of sonodynamic damage to BSA molecules was mainly mediated by singlet oxygen (¹O₂). Otherwise, the binding and damaging sites to BSA molecules were also estimated by synchronous fluorescence. The results indicated that the NR is more vicinal to tryptophan (Trp) residue than to tyrosine (Tyr) residue and the damage site is also mainly at Trp residues. The research result will bring a certain significance to use sonosensitive drugs in the fields of tumor treatment.     

Interaction between titanium dioxide nanoparticles and human serum albumin revealed by fluorescence spectroscopy in the absence of photoactivation

Titanium dioxide (TiO₂) nanoparticles (NPs) are widely used as an important kind of biomaterials. The interaction between TiO₂ (P25) at 20 nm in diameter and human serum albumin (HSA) was studied by fluorescence spectroscopy in this work. Under the simulative physiological conditions, fluorescence data revealed the presence of a single class of binding site on HSA and its binding constants (K_a) were 2.18±0.04×10⁴, 0.87±0.05×10⁴, 0.68±0.06×10⁴ M⁻¹ at 298, 304 and 310 K, respectively. In addition, according to the Van’t Hoff equation, the thermodynamic functions standard enthalpy (ΔH⁰) and standard entropy (ΔS⁰) for the reaction were calculated to be −75.18±0.15 kJ mol⁻¹ and −170.11±0.38 J mol⁻¹ K⁻¹. These results indicated that TiO₂ NPs bond to HSA mainly by van der Waals force and hydrogen bonding formation in low dielectric media, and the electrostatic interactions cannot be excluded. Furthermore, the effects of common ions on the binding constant of TiO₂ NPs-HSA complex were discussed.     

Mechanistic and conformational studies on the interaction of anti-inflammatory drugs, isoxicam and tenoxicam with bovine serum albumin

The mechanism of interaction of the non-steroidal anti-inflammatory drugs, isoxicam (IXM) and tenoxicam (TXM) with bovine serum albumin (BSA) has been studied using spectroscopic techniques, viz., spectrofluorescence, circular dichroism (CD), UV-visible absorption and FT-IR under simulative physiological conditions. Stern-Volmer analysis of fluorescence quenching data shows the presence of the static quenching mechanism. Thermodynamic parameters (negative ΔH⁰ and positive ΔS⁰ values obtained in the present study) revealed that the hydrophobic interactions played a major role in the interaction of these drugs with BSA. The distance, r between the donor (BSA) and acceptor (IXM/TXM) was calculated based on the Forster’s theory of non-radiation energy transfer and the values were observed to be 3.85 nm and 2.60 nm in IXM-BSA and TXM-BSA system, respectively. CD and FT-IR studies indicated that the binding of IXM/TXM to BSA induced conformational changes in BSA. The effect of common ions on the binding of IXM/TXM to BSA has been investigated.     

Study on the Interaction between Florasulam and Bovine Serum Albumin

In this paper, the interaction between florasulam (FU, 2′,6′,8-trifluoro-5-methoxy [Kragh-Hansen U, Molecular aspects of ligand binding to serum albumin. Pharmacol Rev 33(1):17–53 1981; Carter DC and Ho JX, Structure of serum albumin. Adv Protein Chem 45:153–203 1994; He XM, and Carter DC, Atomic structure and chemistry of human serum albumin. Nature 358(6383):209–215 1992] triazolo [1,5-c]pyrimidine-2-sulfonanilide) and bovine serum albumin (BSA) was investigated by fluorescence, ultraviolet absorption (UV) and Far-UV circular dichroism (CD) spectrometries. A strong fluorescence quenching was observed and the quenching mechanism was considered as static quenching. The binding constant of FU with BSA at 299 and 309 K were obtained as 1.5?×?10⁴ and 7.1?×?10³ l mol^?1, respectively. There was one binding site between FU and BSA. The thermodynamic parameters enthalpy change (ΔH) and entropy change (ΔS) were calculated as ?57.89 kJ mol^?1 and ?113.6 J mol^?1 K^?1, respectively, which indicated that the acting force between FU and BSA was mainly hydrogen bond and Van der Waals force. According to the Förster non-radiation energy transfer theory, the average binding distance between donor (BSA) and acceptor (FU) was obtained (r?=?1.59 nm). The investigations of the UV/Vis and CD spectra of the system showed that the conformation of BSA was changed in presence of FU.     

Spectroscopic study on the interaction of Bacillus subtilis α-amylase with cetyltrimethylammonium bromide

The interaction between α‐amylase from Bacillus subtilis and cetyltrimethylammonium bromide (CTAB) has been investigated at various temperature conditions using fluorescence and circular dichroism (CD) spectroscopic methods. Fluorescence data revealed that the fluorescence quenching of α‐amylase by CTAB is the result of complex formation between CTAB and α‐amylase. The thermodynamic analysis on the binding interaction data shows that the interactions are strongly exothermic (ΔH°=−17.92 kJ mol⁻¹) accompanied with increase in entropy (ΔS° between 109 to 135 J mol⁻¹ K⁻¹). Thus the binding of CTAB to α-amylase is both enthalpic and entropic driven, which represent the predominate role of both electrostatic and hydrophobic interactions in complex formation process. The values of 2.17×10⁻³ M⁻¹ and 1.30 have been obtained from associative binding constant (K_a) and stoichiometry of binding number (n), from analysis of fluorescence data, respectively. Circular dichroism spectra showed the substantial conformational changes in secondary structure of α‐amylase due to binding of CTAB, which represents the complete destruction of both secondary and tertiary structure of α-amylase by CTAB.     

Binding of rare earth metal complexes with an ofloxacin derivative to bovine serum albumin and its effect on the conformation of protein

The water-soluble Pr (Ⅲ) and Nd (Ⅲ) complexes with an ofloxacin derivative have been prepared and characterized. The single-crystal X-ray diffraction showed that the Pr (III) and Nd (III) complexes have the similar molecular structure. Under physiological pH condition, the effects of [PrL(NO₃)₂(CH₃OH)](NO₃) and [NdL(NO₃)₂(CH₃OH)](NO₃) on bovine serum albumin (BSA) were examined using fluorescence spectroscopy in combination with UV-vis absorbance and circular dichroism (CD) spectra. The result reveals that the quenching mechanism of fluorescence of BSA by two complexes is a static quenching process and the number of binding sites is about 1 for both. The thermodynamic parameters (ΔH=−14.52 kJ mol⁻¹, ΔS=56.54 J mol⁻¹ K⁻¹ for [PrL(NO₃)₂(CH₃OH)](NO₃) and ΔH=−24.63 kJ mol⁻¹, ΔS=22.07 J mol⁻¹ K⁻¹ for [NdL(NO₃)₂(CH₃OH)](NO₃)) indicate that hydrophobic and electrostatic interactions are the main binding force in the complexes-BSA system. The binding average distance between complexes and BSA was obtained on the basis of Förster's theory. In addition, it was proved by the CD spectra that the BSA secondary structure was changed in the presence of complexes in an aqueous solution.     

Interaction of nobiletin with human serum albumin studied using optical spectroscopy and molecular modeling methods

The binding of nobiletin to human serum albumin (HSA) was investigated by fluorescence, UV-vis, FT-IR, CD, and molecular modeling. Fluorescence data revealed the presence of a single class of binding site on HSA and its binding constants (K) at four different temperatures (289, 296, 303 and 310 K) were 4.054, 4.769, 5.646 and 7.044×10⁴ M⁻¹, respectively. The enthalpy change (ΔH⁰) and the entropy changes (ΔS⁰) were calculated to be 1.938 kJ mol⁻¹ and 155.195 J mol⁻¹ K⁻¹ according to the Van’t Hoff equation. The binding average distance, r, between the donor (HSA) and the acceptor (nobiletin) was evaluated and found to be 2.33 nm according to the Förster's theory of non-radiation energy transfer. Changes in the CD and FT-IR spectra were observed upon ligand binding along with a significant degree of tryptophan fluorescence quenching on complex formation. Computational mapping of the possible binding sites of nobiletin revealed the molecule to be bound in the large hydrophobic cavity of subdomain IIA.     

Spectroscopic Studies on the Interaction of Acid Yellow With Bovine Serum Albumin

Azo dyes, which are common in the environment, can be toxic to various organisms. In order to determine the molecular mechanism of acid yellow 11(AY) toxicity, we studied the effect of AY exposure to the common protein bovine serum albumin (BSA) by several spectroscopic techniques including fluorescence spectroscopy, ultraviolet spectrophotometry (UV) and circular dichroism (CD). It could be concluded from the fluorescence spectra that the quenching effect of BSA by AY was mainly due to complex formation which was unrelated to the absorption of AY. The enthalpy change (ΔH) and entropy change (ΔS) were found to be −21.94 kJ/mol and 30.04 Jmol^-1 K^-1, respectively. The results confirm that electrostatic attraction was the predominant intermolecular force between BSA and AY. Furthermore, the binding distance (r) between AY and the inner tryptophan residue of BSA was determined to be 3.541 nm on the basis of Forster theory of non-radiative energy transfer. In addition, the conformational changes of BSA in the presence of AY were also analyzed by UV and CD. These results indicated that AY could interact with BSA by complex formation, which also affected the structure of BSA.     

Characterize the interaction between naringenin and bovine serum albumin using spectroscopic approach

Naringenin, a flavanone compound highly enriched in grapefruits, has been identified as a possible inhibitor of cell proliferation; and thus has the potential to act as an antitumorigenic agent. In this study, the binding of naringenin to bovine serum albumin (BSA) was studied at the physiological conditions (pH=7.40) by fluorescence and UV-vis spectroscopy. Naringenin strongly quenches the intrinsic fluorescence of BSA, and a decrease in the fluorescence quenching constant was observed together with an increase in temperature, which indicates that the fluorescence quenching of BSA by naringenin is a result of the formation of naringenin-BSA complex. Binding parameters calculating from Stern-Volmer method and Scatchard method showed that naringenin bind to BSA with the binding affinities of the order 10⁴ L mol⁻¹. Thermodynamic parameters such as ΔG, ΔH and ΔS, were calculated at different temperatures, showing that electrostatic interactions were mostly responsible for the binding of naringenin to BSA. Site marker competitive displacement experiments demonstrating that naringenin bind with high affinity to site I (subdomain IIA) of BSA. Furthermore, the effect of metal ions to naringenin-BSA system was studied, and the specific binding distance r (3.30 nm) between donor (Trp-212) and acceptor (naringenin) was obtained according to fluorescence resonance energy transfer (FRET).     

Study of the interaction between fluoxetine hydrochloride and bovine serum albumin in the imitated physiological conditions by multi-spectroscopic methods

The mechanism of interaction of an antidepressant, fluoxetine hydrochloride (FLX) with bovine serum albumin (BSA) has been studied by different spectroscopic techniques under physiological conditions. FLX was found to quench the intrinsic fluorescence of protein by static quenching mechanism. The binding constant ‘K’ was found to be 7.06×10³ M⁻¹ at 296 K. The value of ‘n’ close to unity revealed that the BSA has a single class of binding site for FLX. Based on thermodynamic parameters, hydrogen bonding and van der Waals forces were proposed to operate between BSA and FLX. The change in conformation of protein was noticed upon its interaction with the drug. From displacement studies it was concluded that the FLX bound to protein at site I. The effects of various common metals ions on the binding were also investigated.     

The investigation of the interaction between edaravone and bovine serum albumin by spectroscopic approaches

The fluorescence and ultraviolet spectroscopies were explored to study the interaction between edaravone (EDA) and bovine serum albumin (BSA) under imitated physiological condition. The experimental results show that the fluorescence quenching mechanism between EDA and BSA is a combined quenching (dynamic and static quenching). The binding constants, binding sites, and the corresponding thermodynamic parameters (ΔG, ΔH, and ΔS) of the interaction system were calculated at different temperatures. According to Förster non-radiation energy transfer theory, the binding distance between EDA and BSA was calculated to be 3.10 nm. The effect of EDA on the conformation of BSA was analyzed using synchronous fluorescence spectroscopy. In addition, the effects of some common metal ions Mg²⁺, Ca²⁺, Cu²⁺, and Ni²⁺ on the binding constant between EDA and BSA were examined.     

Static quenching of bovine serum albumin conjugated with small size CdS nanocrystalline quantum dots

The author reports on a strong fluorescence quenching of a model transport protein, bovine serum albumin BSA, when bioconjugated with CdS quantum dots QDs. The 4.4 nm size CdS QDs were synthesized using wet chemistry method and were characterized using UV-vis spectroscopy, scanning electron microscopy SEM and X-ray diffraction XRD techniques. It was found that the BSA fluorescence quenching increases linearly with increasing the CdS QDs concentrations in the range of 3×10⁻⁷-2.0×10⁻⁶ mol L⁻¹. This quenching is explained in terms of Stern-Volmer equation and is ascribed to static quenching with quenching constant 1.321×10⁴ L mol⁻¹ at 300 K.