Spectroscopic studies of interaction between CuO nanoparticles and bovine serum albumin

Recently, the great interests in manufacturing and application of metal oxide nanoparticles in commercial and industrial products have led to focus on the potential impact of these particles on biomacromolecules. In the present study, the interaction of copper oxide (CuO) nanoparticles with bovine serum albumin (BSA) was studied by spectroscopic techniques. The zeta potential value for BSA and CuO nanoparticles with average diameter of around 50 nm at concentration of 10 μM in the deionized (DI) water were ?5.8 and ?22.5 mV, respectively. Circular dichroism studies did not show any changes in the content of secondary structure of the protein after CuO nanoparticles interaction. Fluorescence data revealed that the fluorescence quenching of BSA by CuO nanoparticles was the result of the formed complex of CuO nanoparticles – BSA. Binding constants and other thermodynamic parameters were determined at three different temperatures. The hydrogen bond interactions are the predominant intermolecular forces to stabilize the CuO nanoparticle – BSA complex. This study provides important insight into the interaction of CuO nanoparticles with proteins, which may be of importance for further application of these nanoparticles in biomedical applications.     

Synthesis and application of thiol‐functionalized magnetic nanoparticles for studying interactions of epirubicin hydrochloride with bovine serum albumin by fluorescence spectrometry

A novel method was developed for studying the interaction between epirubicin hydrochloride (EPI) and bovine serum albumin (BSA) by fluorescence spectrometry. Fe₃O₄ magnetic nanoparticles (MNPs) synthesized and functionalized with thiol group were employed for the immobilization and separation of target BSA in reaction solutions. The concentrations of the non‐immobilized BSA and unbound EPI were obtained separately by fluorescence spectrometry. The binding constants (K _a ) and number of binding sites (n ) of EPI with BSA were calculated. In this study, the K _a value was 5.05 × 10⁵ L mol^?1, suggesting a strong binding of EPI to BSA, and the n value was 1.15. The effects of common metal ions on K _a of EPI with BSA were also investigated, and the results showed there was clearly bindings between the metal ions and BSA. The precise binding sites of EPI on BSA were determined as being in site I from the competitive displacement experiments. Copyright © 2016 John Wiley & Sons, Ltd.     

Interaction of Prussian blue nanoparticles with bovine serum albumin: a multi-spectroscopic approach

Owning to their exceptional properties, Prussian blue nanoparticles (PBNPs) are promising in a variety of biomedical applications. In this scenario, understanding of how PBNPs interact and behave in biological systems is essential. Herein, the interaction of PBNPs with protein was investigated. Specifically, the citric acid stabilized PBNPs with a size of 10 nm were synthesized and characterized. The interactions of these PBNPs with the model protein, bovine serum albumin (BSA), were then probed by spectroscopic methods. It was found that the BSA intrinsic fluorescence was quenched upon addition of PBNPs due to the static interaction, suggesting the binding of PBNPs with BSA. Moreover, the synchronous fluorescence and circular dichroism spectra indicated the conformational change of BSA due to the presence of PBNPs.     

Quasi‐spherical silver nanoparticles with high dispersity and uniform sizes: preparation,characterization and bioactivity in their interaction with bovine serum albumin

A stepwise seeded growth route for the preparation of silver nanoparticles (AgNPs) is reported. In the process, silver nitrate was used as a precursor, with sodium borohydride as a reducing agent and trisodium citrate as both a reductant and stabilizer. The AgNPs were characterized using several methods, including UV–vis spectroscopy, X‐ray diffraction and transmission electron microscopy. The prepared AgNPs were quasi‐spherical and crystalline, with an average diameter of 21 nm. Interactions between the AgNPs and bovine serum albumin (BSA) were investigated using UV–vis, fluorescence spectroscopy and synchronous fluorescence spectroscopy (SFS). It was proved that the quenching mechanism is a static process. The binding constants and number of binding sites were calculated. The thermodynamic parameters implied that the binding process was spontaneous and the main driving force of the interaction was electrostatic. The results of the SFS indicated that the conformational change of BSA was induced by AgNPs. Copyright © 2016 John Wiley & Sons, Ltd.     

Conjugate of bovine serum albumin with nicotine

The preparation and characterization of a nicotine-albumin conjugate are reported. The nicotine derivative, 6-(p-aminobenzamido)nicotine, which was coupled to bovine serum albumin(BSA), was synthesized by the following sequence; $\text{1}$-nicotine→6-aminonicotine→6-(p-nitrobenzamido)nicotine→6-(p-aminobenzamido)nicotine. Thirty molecules of conjugated nicotine per molecule of BSA were distinguished by the determination of 6-aminonicotine after hydrolysis. The distribution of the conjugated nicotine on BSA was examined by the stoichiometry of the composed amino acids before and after conjugation, and also by the electrophoretic behavior of the conjugated BSA on a polyacrylamide gel.     

Interaction of wogonin with bovine serum albumin

The binding of wogonin with bovine serum albumin (BSA) was investigated at different temperatures by fluorescence, circular dichroism (CD) and Fourier transform infrared spectroscopy (FT-IR) at pH7.40. The association constants K were determined by Stern-Volmer equation based on the quenching of the fluorescence of BSA in the presence of wogonin, which were in agreement with the constants calculated by Scatchard plots. The thermodynamic parameters were calculated according to the Van't Hoff equation and the result indicated that DeltaH(0) and DeltaS(0) had a negative value (-12.02 kJ/mol) and a positive value (58.72 J/mol K), respectively. On the basis of the displacement experimental and the thermodynamic results, it is considered that wogonin binds to site I (subdomain IIA) of BSA mainly by hydrophobic interaction. The studied results by FT-IR and CD experiment indicated that the secondary structures of protein have been perturbed by the interaction of wogonin with BSA.     

Interactions of cannabinoids with bovine serum albumin

There is no shift of emission maximum (F₄₇₀nm) of bovine serum albumin (BSA)-l-anilino-8-naphthatene sulphonic acid (ANS) complex in the pesence of delta-9-tetrahydrocannabinol (delta-9-THC) alone and cannabidiol (CBD) or cannabinol (CBN) in the presence and absence of delta-9-THC. Delta-9-THC (1.66–13.33 M) and CBD at higher concentrations (13.33–20.0 M) produce a concentration-dependent significant quenching of fluorescence of BSA-ANS complex, but CBN (l.66–20.0 M) as well as CBD at lower concentrations (1.66–6.66 M) fails to produce any significant change in Iluorescence intensity under similar conditions. Furthermore, neither CBD nor CBN is able to affect the delta-9-THC-induced quenching of fluorescence intensity of BSA-ANS complex. These results indicate that the binding of cannabinoids to the ANS binding sites of BSA molecule are in the order detta-9-THC > CBr3 > CBN, and CBD or CBN does not have any influence on the binding of delta-9-THC to BSA molecules under these conditions.     

Effects of metallic silver particles on resonance energy transfer in labeled bovine serum albumin

Resonance energy transfer (RET) is widely used to detect proximity between biomolecules. In transparent solution the maximum donor-to-acceptor distance for RET is about 70 A. We measured the effects of metallic silver island films on RET from the intrinsic tryptophan of a protein to a bound probe as the acceptor. These preliminary experiments revealed a dramatic increase in the apparent F?rster distance increasing from 28.6 to 63 A. These results suggest the use of silver island films for detecting long range proximity between biomolecules and for biotechnology applications based on RET.     

Interaction of ergosterol with bovine serum albumin and human serum albumin by spectroscopic analysis

This study was designed to examine the interactions of ergosterol with bovine serum albumin (BSA) and human serum albumin (HSA) under physiological conditions with the drug concentrations in the range of 2.99-105.88?μM and the concentration of proteins was fixed at 5.0?μM. The analysis of emission spectra quenching at different temperatures revealed that the quenching mechanism of HSA/BSA by ergosterol was the static quenching. The number of binding sites n and the binding constants K were obtained at various temperatures. The distance r between ergosterol and HSA/BSA was evaluated according to F?ster non-radioactive energy transfer theory. The results of synchronous fluorescence, 3D fluorescence, FT-IR, CD and UV-Vis absorption spectra showed that the conformations of HSA/BSA altered in the presence of ergosterol. The thermodynamic parameters, free energy change (ΔG), enthalpy change (ΔH) and entropy change (ΔS) for BSA-ergosterol and HSA-ergosterol systems were calculated by the van't Hoff equation and discussed. Besides, with the aid of three site markers (for example, phenylbutazone, ibuprofen and digitoxin), we have reported that ergosterol primarily binds to the tryptophan residues of BSA/HSA within site I (subdomain II A).     

Interaction of allylisothiocyanate with bovine serum albumin

The interaction of allylisothiocyanate with bovine serum albumin was monitored by fluorescence titration. The interaction was weak with an apparent association constant of 2 × 10². The interaction was unaffected in the pH range of 5.0 to 8.3 and by NaCl. However, the addition of dioxane upto 4% increased the value of the association constant. N-Methyl bovine serum albumin and bovine serum albumin with sulphydryl groups blocked had the same affinity for allylisothiocyanate suggesting that amino and sulphydryl groups may not be involved in the interaction. Polyacrylamide gel electrophoresis and estimation of available lysine suggested that there were perhaps two types of groups involved in the interaction of allylisothiocyanate with bovine serum albumin. An erratum to this article is available at .     

Interaction of new pyridylporphyrins with bovine serum albumin

The interaction of meso-tetra(4-N-hydroxyethylpyridyl)porphyrin, meso-tetra(3-N-hydroxyethylpyridyl)porphyrin, and their zinc complexes with bovine serum albumin (BSA) was studied by electronic spectroscopy, CD, and equilibrium dialysis at pH 7.2. The titration of the porphyrins with BSA was accompanied by a decrease in light absorption and a bathochromic shift of the Soret band, as well as by the appearance of an isobestic point. The porphyrin interaction with BSA also led to the induction of positive CD spectra in the visible region, which is explained by the porphyrin sorption on the protein globule. The equilibrium dialysis helped in determining the stoichiometry of binding and the binding constants of the porphyrins under study with BSA using Scatchard plots. This interaction is nonspecific and reversible. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 2; see also http://www.maik.ru.     

Study on the antibacterial activity and interaction with human serum albumin of Tagetes erecta inspired biogenic silver nanoparticles

Here, Tagetes erecta leaves extract mediated silver nanopartices (te-SNPs) were synthesized. This synthesis process was simple, one step and eco-friendly. The te-SNPs were characterized by various spectroscopic instruments such as ultraviolet visible (UV–vis), Fourier-transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS) and Transmission Electron Microscopy (TEM) with energy Dispersive X-ray Spectroscopy (EDS). These te-SNPs showed the effective bactericidal activity against Escherichia coli and Staphylococcus aureus. The interaction of te-SNPs with human serum albumin (HSA) was evaluated by UV–vis absorption, fluorescence (FL), time resolved fluorescence (TRF), DLS and circular dichroism (CD) study. TRF and temperature dependant study suggested that the interaction process followed the dynamic quenching mechanism. The stern-volmer quenching constants (K_SV) were obtained as 1.36 × 10⁷, 1.51 × 10⁷ and 1.94 × 10⁷ M⁻¹at 288, 298 and 308 K respectively. The thermodynamic parameters were also evaluated and this result suggested that the interaction was spontaneous, and the hydrophobic forces played the most important role for the interaction process. In addition, CD spectra proved minute alteration of secondary structure of HSA upon interaction with the te-SNPs.     

Comparison of interactions between human serum albumin and silver nanoparticles of different sizes using spectroscopic methods

Three different sizes (15.9 ± 2.1 nm, 26.4 ± 3.2 nm and 39.8 ± 4.0 nm, respectively) of citrate‐coated silver nanoparticles (SNPs) have been synthesized and characterized. The interactions of the synthesized SNPs with human serum albumin (HSA) at physiological pH have been systematically studied by UV‐vis absorption spectroscopy, fluorescence spectroscopy, synchronous fluorescence spectroscopy, three‐dimensional fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The results indicate that the SNPs can bind to HSA with high affinity and quench the intrinsic fluorescence of HSA. The binding constants and quenching rate constants were calculated. The apparent association constants (K_app) values are 2.14 × 10⁴ M^–1 for 15.9 nm SNP, 1.65 × 10⁴ M^–1 for 26.4 nm SNP and 1.37 × 10⁴ M^–1 for 39.8 nm SNP, respectively. The values of binding constant obtained from the fluorescence quenching data match well with that determined from the absorption spectral changes. These results suggest that the smaller SNPs have stronger interactions to HSA than the larger ones at the same concentrations. Synchronous fluorescence, three‐dimensional fluorescence and CD spectroscopy studies show that the synthesized SNPs can induce slight conformational changes in HSA. Copyright © 2014 John Wiley & Sons, Ltd.     

Aggregation and fibrillation of bovine serum albumin

The all-alpha helix multi-domain protein bovine serum albumin (BSA) aggregates at elevated temperatures. Here we show that these thermal aggregates have amyloid properties. They bind the fibril-specific dyes Thioflavin T and Congo Red, show elongated although somewhat worm-like morphology and characteristic amyloid X-ray fiber diffraction peaks. Fibrillation occurs over minutes to hours without a lag phase, is independent of seeding and shows only moderate concentration dependence, suggesting intramolecular aggregation nuclei. Nevertheless, multi-exponential increases in dye-binding signal and changes in morphology suggest the existence of different aggregate species. Although beta-sheet content increases from 0 to ca. 40% upon aggregation, the aggregates retain significant amounts of alpha-helix structure, and lack a protease-resistant core. Thus BSA is able to form well-ordered beta-sheet rich aggregates which nevertheless do not possess the same structural rigidity as classical fibrils. The aggregates do not permeabilize synthetic membranes and are not cytotoxic. The ease with which a multidomain all-alpha helix protein can form higher-order beta-sheet structure, while retaining significant amounts of alpha-helix, highlights the universality of the fibrillation mechanism. However, the presence of non-beta-sheet structure may influence the final fibrillar structure and could be a key component in aggregated BSA's lack of cytotoxicity.     

Thermal aggregation of bovine serum albumin at different pH: comparison with human serum albumin

We report here a study on thermal aggregation of BSA at two different pH values selected to be close to the isoelectric point (pI) of this protein. Our aim is to better understand the several steps and mechanisms accompanying the aggregation process. For this purpose we have performed kinetics of integrated intensity emission of intrinsic and extrinsic dyes, tryptophans and ANS respectively, kinetics of Rayleigh scattering and of turbidity. The results confirm the important role played by conformational changes in the tertiary structure, especially in the exposure of internal hydrophobic regions that promote intermolecular interactions. We also confirm that the absence of electrostatic repulsion favours the disordered non-specific interactions between molecules and consequently affects the aggregation rate. Finally, the comparison between BSA and another relative protein, HSA, allows us to clarify the role of different domains involved in the aggregation process. Proceedings of the XVIII Congress of the Italian Society of Pure and Applied Biophysics (SIBPA), Palermo, Sicily, September 2006.