Conformation Of BSA Research Articles

Enhanced enzymatic activity and conformational stability of catalase in presence of tetrahedral DNA nanostructures: A biophysical and kinetic study

The emergence of DNA nanotechnology has shown enormous potential in a vast array of applications, particularly in the medicinal and theranostics fields. Nevertheless, the knowledge on the biocompatibility between DNA nanostructures and cellular proteins is largely unknown. Herein, we report the biophysical interaction between proteins (circulatory protein bovine serum albumin, BSA, and the cellular enzyme bovine liver catalase, BLC) and tetrahedral DNA (tDNA), which are well-known nanocarriers for therapeutics. Interestingly, the secondary conformation of BSA or BLC was unaltered in the presence of tDNAs which supports the biocompatible property of tDNA. In addition, thermodynamic studies showed that the binding of tDNAs with BLC has a stable non-covalent interaction via hydrogen bond and van der Waals contact, which is indicative of a spontaneous reaction. Furthermore, the catalytic activity of BLC was increased in the presence of tDNAs after 24 h of incubation. These findings indicate that the presence of tDNA nanostructures not only ensures a steady secondary conformation of proteins, but also stabilize the intracellular proteins like BLC. Surprisingly, our investigation discovered that tDNAs have no effect on albumin proteins, either by interfering or by adhering to the extracellular proteins. These findings will aid in the design of future DNA nanostructures for biomedical applications by increasing the knowledge on the biocompatible interaction of tDNAs with biomacromolecules.

Enantioselective effect of chiral prothioconazole on the conformation of bovine serum albumin

As a typical chiral triazole fungicide, the enantioselective toxicity of prothioconazole to environmental organisms is of increasing concern. Herein, the binding mechanism of chiral PTCs to BSA was investigated by multi-spectral technique and molecular docking. Fluorescence titration and fluorescence lifetime experiments fully established that quenching BSA fluorescence by chiral PTCs is static quenching and could spontaneously bind to BSA. Hydrophobic interactions dominate the binding process of chiral PTCs to BSA. Differently, although both chiral PTCs and BSA have a primary binding site, the difference in chiral isomerism leads to a stronger binding ability of S-PTC than R-PTC. Both configurations of PTC can change the conformation of BSA and induce changes in the microenvironment around its amino acid residues, and the effect of S-PTC is more significant. Overall, S-PTC exhibited a more substantial effect on BSA structure relative to R-PTC. That is, S-PTC may lead to more potent potential toxicological effects on environmental organisms. This study provides a comprehensive assessment of the environmental behavior of chiral pesticides and their potential toxicity to environmental organisms at the molecular level and provides a theoretical basis for the screening of highly effective and biologically less toxic enantiomers of chiral pesticides.

A Photo-Responsive Porphyrin-Mn@Choles Complex for Bacteria Treatment

Biocompatible photo-driven producers of singlet oxygen can inhibit the growth of drug-resistant bacteria and tumors. In order to develop bacteria targeting generator of singlet oxygen for tumor and bacterial treatment, a metal porphyrin liposome (Phy–Mn–Ls) was prepared by the metal coordination reaction and self-assembly of porphyrin compounds with bacteria targeting polymer (HS–PEG–chol). The photo-driven production of 1O2, binding with bovine serum protein (BSA) and lipase, toxicity to MCF-7 breast cancer cells and inhibitory effect on the growth of Escherichia coli have been investigated. Fluorescence analysis results show that Phy–Mn–Ls can bind to lipase, and it shows less effect on the conformation of BSA and is low cytotoxicity without irradiation. In particular, the good biocompatibility made Phy–Mn–Ls exhibit good photosensitive antibacterial activity and anti-tumor properties. The results demonstrate that the coordination of HS–PEG–chol with metal-phorphrin coodination is an effective way to develop bacteria targeting nano-complexes (Phy–Mn–Ls) for lipase affinity and photodriven bacteria treatment.

Interaction study of monoisoamyl dimercaptosuccinic acid with bovine serum albumin using biophysical and molecular docking approaches

Monoisoamyl 2,3-dimercaptosuccinic acid (MiADMSA), a lipophilic chelator has been evaluated for its potential use as an antidote in arsenic poisoning. The pharmacokinetics and pharmacodynamics properties of a drug could be understood via study its mechanism of interaction with bovine serum albumin protein (BSA). Therefore, the interaction between MiADMSA with BSA was investigated using various spectroscopic techniques and computational methods. Linear quenching of BSA intrinsic fluorescence intensity with the increasing concentration of MiADMSA was observed in the fluorescence study. Furthermore, synchronous results revealed that MiADMSA slightly changed the conformation of BSA. The binding constant value of the BSA-MiADMSA complex was found 1.60 × 104 M−1 at 298 K. The value of thermodynamic parameters ΔG, ΔH, and ΔS described that the process is spontaneous, endothermic, and hydrophobic forces are involved in the interaction of MiADMSA with BSA. Competitive site marker experiments showed that MiADMSA binds to site-II of BSA. Conformational changes of BSA with the interaction of MiADMSA were apparent by CD, UV–Visible, FT-IR, and 3D fluorescence spectroscopy. To strengthen the experimental findings we have also performed a theoretical study on the BSA-MiADMSA complex. Two sites were identified with docking score of − 6.642 kcal/mol at site IIa and − 3.80 kcal/mol for site IIb via molecular docking study. Molecular dynamics simulation study inferred the stability of the BSA-MiADMSA complex which was analyzed in a long simulation run. The experimental and computational studies have shown the effective binding of MiADMSA with BSA which is essential for the transportation and elimination of a drug from the body.

A potent drug candidature of Cu(II) pyrazino[2,3‐f][1,10]phenanthroline complexes with bioactive ligands: synthesis, crystal structures, biomolecular interactions, radical scavenging and cytotoxicities

A novel ternary copper(II) complexes, - [Cu(py-phen)(asn)(NO3)(H2O)] (1) and [Cu(py-phen)(trp)(H2O)]NO3 (2)- (py-phen: pyrazino[2,3‐f][1,10]phenanthroline, asn: asparagine, trp: tryptophan), have been synthesized and characterized by CHN analysis, ESI-MS, FTIR and single-crystal X-ray diffraction techniques. Interaction of the complexes 1 and 2 with CT-DNA has been investigated by absorption spectral titration, EB and Hoechst 33258 displacement assay. The interaction between the complexes 1 and 2 and BSA was investigated by electronic absorption and fluorescence spectroscopy methods. The experimental outcomes indicate that the fluorescence quenching mechanism between the complexes 1 and 2 and BSA is a static quenching process. The Stern-Volmer constants, binding constants, binding sites and the corresponding thermodynamic parameters (ΔG, ΔH, ΔS) of BSA + complex systems were determined at different temperatures. The binding distance between the complexes 1 and 2 and BSA was calculated according to FRET. The effect of the complexes 1 and 2 on the conformation of BSA was also examined using synchronous, two dimensional (2D) and three dimensional (3D) fluorescence spectroscopy. Radical scavenging activity of the complex was determined in terms of EC50, using the DPPH and H2O2 method. The anticancer activities of the complexes 1 and 2 were investigated using an XTT assay against three cancer cell lines (MCF-7, Caco-2 and A549) and non-tumor cell line (BEAS-2B). Communicated by Ramaswamy H. Sarma

Interactions of antinutrients mixtures with bovine serum albumin and its influence on in vitro protein digestibility

Antinutrient mixtures are usually present in food formulations what can affect the protein digestibility. This work aimed to study the interactions of a mixture of antinutrients with bovine serum albumin and its effect on the in vitro protein digestion. Results showed that the phytic and tannic acid decrease the α-helix conformation of BSA and form insoluble complexes. The phytic acid and saponin increase the affinity between the tannic acid and the protein and change the nature of the forces involved in the binding as observed by isothermal titration calorimetry. The effect of each antinutrient on protein properties depends on the concentration and structure of the protein and the antinutrient. The tannic acid was the antinutrient that most decreased the protein digestion. The presence of both phytic acid and saponin changed the digestibility of protein containing tannin. The study of a mixture of substances is complex and involves several mechanisms of binding and molecules structure changes. This works shows that, despite the complexity of the system studied, the balance between the proportions of each component can determine its influence on the bioavailability of proteins and antinutrients. Thus, the results obtained in the present work may help in the understanding of reactions and protein behavior of complex mixtures such as food matrixes.

An insight into the inhibition of fibrillation process verses disaggregation of preformed fibrils of bovine serum albumin by isoprenaline hydrochloride

This study is based on the analysis of the recent trend of medication in neurodegenerative diseases. Due to the asymptomatic nature of the diseases, medication delays. Therefore, mechanism of medication assists in removal of the symptoms. Therefore, in order to find out remedy for complete prevention of the disease we have considered “inhibition verses disaggregation” study. Various biophysical techniques such as turbidity measurement (TM), Thioflavin T (ThT) binding assays, circular dichroism (CD), transmission electron microscopy (TEM) etc. has been performed. Isoprenaline hydrochloride (ISO) was a good candidate for inhibition and disaggregation of preformed fibrils of BSA. Therefore, it is concluded that inhibition of fibrillation process was more momentous, effective procedure in restricting the aggregation by stabilizing the native conformation of BSA than the removal of preformed amyloid fibrils under in vitro condition. Forwarding ahead, to understand the efficiency of the two processes under in vivo condition, this study can be applied on animal models so that we can look forward on human beings as well for the development of vaccines. This study is concerned about the applied aspect of research in future so that we can hope for prevention of the disease instead of only removal of the symptoms.

Different effects of Forsythia suspensa metabolites on bovine serum albumin (BSA).

Forsythia suspensa metabolites have many bioactivities, such as selective immuno suppression, antioxidation, anti-hepatic injury, etc. In the present study, the interactions of the three metabolites with BSA have been investigated in a buffer (pH 7.40) using multi-spectroscopic techniques in combination with molecular docking methods. Two isoformers, forsythoside A and forsythoside I can statically quench BSA intrinsic fluorescence by forming the complexes with BSA at stoichiometric ratio of 1:1 that is again proved by UV-visible absorption. During the binding, the proportion of α-helix in BSA increases, the microenvironment around Tryptophan 213 changes and FRET is one of the major factors to quench fluorescence. Forsythoside E forms BSA-forsythoside E complex (1:1) and thus enhances the intrinsic fluorescence of BSA. During the process, forsythoside E affects not only Tryptophan residues but also Tyrosine residues so that the conformation of BSA is consequently changed. All above binding processes are spontaneous mainly through hydrogen bonding and the hydrophobic force interaction, which is supported by docking analysis and thermodynamic parameters. In addition, three compounds do not induce BSA aggregation. These findings are beneficial to understand the detailed information of the interactions of Forsythia suspensa metabolites with BSA.

Luminophore Formation in Various Conformations of Bovine Serum Albumin by Binding of Gold(III).

The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (λem = 640 nm) of BSA-Au(III) complexes. The method adjusts the pH to show that the emergence of the red fluorescence is correlated with the pH-induced equilibrium transitions of the BSA conformations. Red fluorescent BSA-Au(III) complexes can only be formed with an adjustment of pH at or above 9.7, which corresponds to the "A-form" conformation of BSA. The protocol to adjust the BSA to Au molar ratio and to monitor the time-course of the process of Au(III) binding is described. The minimum number of Au(III) per BSA, to produce the red fluorescence, is less than seven. We describe the protocol in steps to illustrate the presence of multiple Au(III) binding sites in BSA. First, by adding copper (Cu(II)) or nickel (Ni(II)) cations followed by Au(III), this method reveals a binding site for Au(III) that is not the red fluorophore. Second, by modifying BSA by thiol capping agents, another nonfluorophore-forming Au(III) binding site is revealed. Third, changing the BSA conformation by cleaving and capping of the disulfide bonds, the possible Au(III) binding site(s) are illustrated. The protocol described, to control the BSA conformations and Au(III) binding, can be generally applied to study the interactions of other proteins and metal cations.

Insights into the binding behavior of bovine serum albumin to black carbon nanoparticles and induced cytotoxicity

Black carbon (BC) is a main component of particulate matter (PM2.5). Due to their small size (<100nm), inhaled ultrafine BC nanoparticles may penetrate the lung alveoli, where they interact with surfactant proteins and lipids, causing more serious damage to human health. Here, BC was analyzed to investigate the binding mechanism of its interaction with protein and induction of cytotoxicity changes. The binding process and protein conformation between BC and a serum protein (bovine serum albumin, BSA) were monitored by using a fluorescence quenching technique and UV-vis absorption, Fourier transform infrared (FTIR) and circular dichroism (CD) spectroscopies. The experimental results revealed that the fluorescence quenching of BSA induced by BC was a static quenching process and the hydrophobic force played the critical role in the interaction. The native conformation of BSA on the BC surface was slightly disturbed but obvious structural unfolding of the secondary structure did not occur. In the cytotoxicity study, BC nanoparticles with low concentrations exhibited strong toxicity towards BEAS-2B cells. However, the toxicity of BC nanoparticles could be mitigated by the presence of BSA. Therefore, proteins in biological fluids likely reduce the toxic effect of BC on human health. These findings delineated the binding mechanism and the toxicity between BC and the BSA-BC system, contributing to the understanding of the biological effects of BC exposure on human health in polluted atmospheres.

Cyanidin-3-O-glucoside functions like chemical chaperone and attenuates the glycation mediated amyloid formation in albumin

In this study, chemical chaperone like function of cyanidin-3-O-glucoside (C3G) was investigated through fluorescence spectroscopy, UV–visible spectroscopy, circular dichroism spectroscopy, confocal microscopy, scanning electron microscopy and molecular docking studies. Early and advanced glycation inhibitory effect was evaluated by fluorescence spectroscopy and agarose gel electrophoresis. Amyloids were investigated based on their propensity to bind Congo Red (CR) and Thioflavin T (ThT) by multiple microscopic approaches. Circular dichroism studies were used to analyze the changes in the secondary structure due to glycation. C3G effectively inhibited early and advanced glycation by masking like function, carbonyl scavenging and chemical chaperone activity. C3G had molecular interaction with Glu186, Arg427, Ser428, Lys431, Arg435, and Arg458 of BSA. Based on the microscopic analysis, it is evident that C3G can inhibit protein aggregation and amyloid formation. Circular dichroism studies suggested that glycation had resulted in augmented β-sheet propensity, whereas C3G had a protective effect on the helical conformation of BSA. We conclude that C3G has a chemical chaperone like function on the event of glycation mediated amyloid formation in BSA.

Spectrophotometric and molecular modelling studies on in vitro interaction of tyrosine kinase inhibitor linifanib with bovine serum albumin.

Linifanib (LNF) possess antitumor activity and acts by inhibiting receptor tyrosine kinase VEGF and PDGF. The interaction of BSA with the drug can provide valuable information regarding the pharmacokinetic and pharmacodynamics behavior of drug. In our study the spectrophotometric methods and molecular docking studies were executed to understand the interaction behavior of BSA and LNF. BSA has an intrinsic fluorescence and that fluorescence was quenched by LNF. This quenching process was studied at three different temperatures of 288, 300and 308 K. The interaction between LNF and BSA was due to static quenching because the Ksv (Stern-Volmer constant) at 288 K was higher than at 300 and 308 K. Kq (quenching rate constant) behaved in a similar fashion as the Ksv. Several other parameters like binding constants, number of binding sites and binding energy in addition to molecular docking studies were also used to evaluate the interaction process. A decrease in the binding constants was observed with increasing temperatures and the binding site number approximated unity. The decreasing binding constant indicates LNF–BSA complex stability. The site mark competition experiment confirmed the binding site for LNF was located on site II of BSA. UV–visible studies along with synchronous fluorescence confirm a small change in the conformation of BSA upon interaction with LNF. The thermodynamic analysis provided the values for free energy ΔG0, ΔH0 and ΔS0. The ΔG0 at the 288, 300 and 308 K ranged in between -21.5 to -23.3 kJ mol-1, whereas the calculated values of ΔH (-55.91 kJ mol-1) and ΔS0 (-111.74 J mol-1·K-1). The experimental and molecular docking results suggest that the interaction between LNF and BSA was spontaneous and they exhibited hydrogen bonding and van der Waals force between them.

Magnetic solid-phase extraction of protein with deep eutectic solvent immobilized magnetic graphene oxide nanoparticles

As a new type of green solvent, four kinds of choline chloride (ChCl)-based deep eutectic solvents (DESs) have been synthesized, and then a core–shell structure magnetic graphene oxide (Fe3O4–NH2@GO) nanoparticles have been prepared and coated with the ChCl-based DESs. Magnetic solid-phase extraction (MSPE) based Fe3O4–NH2@GO@DES was studied for the first time for the extraction of proteins. The characteristic results of vibrating sample magnetometer (VSM), X-ray diffraction (XRD), Fourier transform infrared spectrometry (FT-IR), thermal gravimetric analysis (TGA) and field emission scanning electron microscopy (FESEM) indicated the successful preparation of Fe3O4–NH2@GO@DES. The concentrations of proteins in studies were determined by a UV–vis spectrophotometer. The advantages of Fe3O4–NH2@GO@DES in protein extraction were compared with Fe3O4–NH2@GO and Fe3O4–NH2, and Fe3O4–NH2@GO@ChCl–glycerol was selected as the suitable extraction solvent. The influence factors of the extraction process such as the pH value, the temperature, the extraction time, the concentration of protein and the amount of Fe3O4–NH2@GO@ChCl–glycerol were evaluated. Desorption experimental result showed 98.73% of BSA could be eluted from the solid extractant with 0.1mol/L Na2HPO4 solution contained 1mol/L NaCl. Besides, the conformation of BSA was not changed during the elution by the investigation of circular dichromism (CD) spectra. Furthermore, the analysis of real sample demonstrated that the prepared magnetic nanoparticles did have extraction ability on proteins in bovine whole blood.

Raman Characterization of Nanoparticle Transport in Microfluidic Paper-Based Analytical Devices (μPADs).

There is great interest in the use of microfluidic paper-based analytical devices (μPADs) for low-cost diagnostics. In this contribution, we illustrate the utility of Raman spectral imaging for both μPAD characterization and for quantification of the transport of applied reagents and analytes within these devices. We evaluated the transport of nanoscale particles within μPADs using a suite of differentially functionalized gold (AuNP) and silver (AgNP) nanoparticles with diameters of 8-64 nm. Nanoparticle transport within the cellulose matrix was characterized by collection of both Raman and surface-enhanced Raman spectroscopy (SERS) spectral maps that enabled differentiation of cellulose fibers and characterization of analyte deposition patterns. The transport of citrate (cit), BSA, PEG, PVP, and DNA functionalized AuNP and AgNP in wax-printed μPADs was primarily affected by nanoparticle surface chemistry rather than particle size or core composition. Sample pH (3-10) influenced the transport of 15 nm BSA-cit-AuNP, but not 15 nm cit-AuNP, because of the effects of solution pH on the charge and conformation of BSA. Derjaguin, Landau, Verwey, and Overbeek theory (DLVO) and extended DLVO (xDLVO) theory are used to explain the collected experimental results.

Spectroscopic investigations on the binding of leucomalachite green to bovine serum album

The interaction of bovine serum with leucomalachite green (LMG) was investigated by spectroscopic methods. LMG was found to quench the intrinsic fluorescence of BSA by dynamic quenching. High quenching constant suggested that LMG can form a complex with BSA. Binding constant and number of binding site were determined. The thermodynamic parameters suggested the main interacting force is hydrophobic forces. The effect of LMG on the conformation of BSA was analyzed by synchronous fluorescence spectroscopy. RLS spectra were also used to reconfirm the formation of complex between BSA and LMG.

Interactions of bovine serum albumin with cationic imidazolium and quaternary ammonium gemini surfactants: Effects of surfactant architecture

The interactions of BSA with a series of cationic imidazolium gemini surfactants ([Cn-s-Cnim]Br2, n=10, 12, 14, s=2, 4, 6), quaternary ammonium surfactants (C12C2C12), and their corresponding monomers ([C12mim]Br and DTAB) are investigated by fluorescence using pyrene as a molecular probe, synchronous fluorescence, circular dichroism (CD), and UV–visible absorption spectra. These surfactants are used to elucidate the effects of surfactant hydrophilic head group, spacer length, and hydrophobic chain length on the conformation of BSA. The results of fluorescence spectra and CD show that the imidazolium gemini surfactants with shorter spacers or with longer hydrophobic chains have a larger effect on BSA unfolding, and the imidazolium gemini surfactant interacts with BSA more strongly than its corresponding monomer and the quaternary ammonium gemini surfactant. These conclusions have been confirmed by the binding constants (Ka) and binding sites (n) for the BSA/surfactant system. Stern–Volmer quenching constants KSV of cationic surfactants binding to BSA are obtained, indicating that the probable quenching mechanism is initiated by ground-state complex formation rather than by dynamic collision. Moreover, the synchronous fluorescence spectra show that the surfactants mainly interact with tryptophan residues of BSA.

Protein Conformational Studies for Macromolecularly Imprinted Polymers

CD is used to clearly show the negative impact of common ligands on the overall conformation of BSA, a typical protein template in macromolecularly imprinted polymers. This change occurs at concentrations far lower than those generally used in the literature. These findings are important as they offer insight into a potential fundamental reason for the lack of success in protein imprinting to date despite significant interest from the scientific community.

Spectroscopic and electrochemical studies on the interaction of an inclusion complex of β-cyclodextrin/fullerene with bovine serum albumin in aqueous solution

Abstract The potential effect of human exposure to carbonaceous nanomaterials (e.g., fullerenes or their derivatives) in the environment has become a concern. In the current study, we report the interaction of one water-soluble fullerene with bovine serum albumin using spectroscopic and electrochemical methods under aqueous solutions. The novel supramolecular inclusion complex of the water-soluble fullerene ( β -CD) 2 /C 60 was synthesized and characterized. In the mechanism discussed, the spectroscopic methods such as fluorescence quenching and ultraviolet–visible absorption, proved that the fluorescence quenching of BSA by ( β -CD) 2 /C 60 was the result of the formation of ( β -CD) 2 /C 60 –BSA complex and that the mechanism of quenching might be a static quenching procedure. The binding constants K a , the number of binding sites n , and the corresponding thermodynamic parameters Δ G , Δ H , and Δ S at different temperatures were calculated through fluorescence spectroscopy, then as an auxiliary method, the electrochemical impedance spectroscopy (EIS) experiments confirmed this conclusion. The results indicated that the electrostatic interactions play a major role in ( β -CD) 2 /C 60 –BSA association. The circular dichroism spectra show the conformation change of the effect of ( β -CD) 2 /C 60 on the conformation of BSA, which was confirmed by the results of the three-dimensional fluorescence spectra. Site marker competitive experiments indicate that the binding of ( β -CD) 2 /C 60 to BSA primarily took place in site I. The distance r between donor (BSA) and acceptor (( β -CD) 2 /C 60 ) was obtained according to fluorescence resonance energy transfer (FRET). This work aims to demonstrate the mechanisms of the formation of the complex between water-soluble fullerene and protein under physiological conditions, as well as the remediation for the possible unwarranted biological effects of water-soluble fullerene.

Study of the Binding of Herbacetin to Bovine Serum Albumin by Fluorescence Spectroscopy

In this paper, the interaction between herbacetin and BSA was investigated by fluorescence and three-dimensional fluorescence spectroscopy under simulated physiological conditions. It was proved that the fluorescence quenching of BSA by herbacetin was mainly the result of the formation of a herbacetin–BSA complex. The modified Stern–Volmer quenching constant and the corresponding thermodynamic parameters ΔH0, ΔG0 and ΔS0 were calculated at different temperatures. The results indicated that electrostatic interactions were the predominant intermolecular forces in stabilizing the complex. The distance r=3.23 nm between the donor (BSA) and acceptor (herbacetin) was obtained according to Forster’s nonradioactive energy transfer theory. The synchronous fluorescence and three-dimensional fluorescence spectra results showed that the hydrophobity of amino acid residues increased in the presence of herbacetin. These results revealed that the microenvironment and conformation of BSA changed during the binding reaction.

Fluorescence studies of interaction between flavonol p-coumaroylglucoside tiliroside and bovine serum albumin

In this paper, the interaction between flavonol p-coumaroylglucoside tiliroside and BSA was investigated by fluorescence quenching spectra, synchronous fluorescence spectra, and three-dimensional fluorescence spectra under simulative physiological conditions. It was proved that the fluorescence quenching of BSA by tiliroside was mainly a result of the formation of a tiliroside–BSA complex. The modified Stern-Volmer quenching constant and the corresponding thermodynamic parameters Δ H, Δ G and Δ S at different temperatures were calculated. The results indicated that electrostatic interactions were the predominant intermolecular forces in stabilizing the complex. The distance r = 3.95 nm between the donor (BSA) and acceptor (tiliroside) was obtained according to Förster's nonradioactive energy transfer theory. The synchronous fluorescence and three-dimensional fluorescence spectra results showed the microenvironment and conformation of BSA were changed in the binding reaction.