Bovine Serum Albumin Complex Research Articles

Complexity of the Role of Various Site-Specific and Selective Sudlow Binding Site Drugs in the Energetics and Stability of the Acridinedione Dye-Bovine Serum Albumin Complex: A Molecular Docking Approach.

Molecular docking (Mol.Doc) techniques were employed to ascertain the binding affinity of two resorcinol-based acridinedione dyes (ADR1 and ADR2) with the widely studied globular protein Bovine Serum Albumin (BSA) in the presence of site-selective binding drugs by Autodock Vina 4.2 software. Docking of various feasible conformers of ADR1 dye with BSA was found to be energetically more favored than ADR2 dye, even though both these dyes differ in the 9th position of the basic dye structure. Analysis of dyes with BSA establishes the location of dye in all of the binding sites of BSA, predominantly through conventional and nonconventional hydrogen-bonding (HB) interactions. The coexistence of hydrophobic interactions resulted in the stability of various conformers generated. The introduction of site I and site II (Sudlow site binding drugs) into ADR1-BSA and ADR2-BSA complexes effectively destabilizes the dye-protein complex; however, the drugs do not displace ADR dyes completely from their selective binding domains. Site II binding drugs effectively destabilize the binding ability of the dye-protein complex rather than site I drugs. However, docking of site I drug 3-carboxyl-4-methyl-5-propyl-2-furanpropanic acid (CMPF) largely destabilizes the ADR1-protein complex, whereas indomethacin (INDO) enhances the binding affinity of the ADR2-protein complex. Interestingly, simultaneous docking of ADR dyes to the BSA-drug complex results in larger stability of the protein-drug complex through HB interactions rather than hydrophobic interactions. Both ADR1 and ADR2 dyes predominantly occupy the Sudlow binding sites of BSA, and the introduction of either site I or site II binding drugs does not displace the dye efficiently from the corresponding binding sites, rather the drugs are effectively displaced toward other binding domains apart from their specific site-binding domains of BSA. Through Mol.Doc techniques, we authenticate that the interactions in host-guest complex systems involving competing ligands are established in depth, wherein the dye as well as the amino acid (AA) moieties in BSA act as both HB donor and acceptor sites apart from several hydrophobic interactions coexisting toward the stability.

Binding of ankaflavin with bovine serum albumin (BSA) in the presence of carrageenan and protective effects of Monascus yellow pigments against oxidative damage to BSA after forming a complex with carrageenan.

Ankaflavin (AK) is a typical yellow pigment extracted from Monascus-fermented rice with several biological effects; however, its solubility is poor. Thus, research studies of the delivery systems of AK, especially those constructed from protein-polysaccharide complexes, have attracted considerable attention. However, the interactions that exist in the system have rarely been investigated. This work focused on the interactions between AK and bovine serum albumin (BSA) as well as the influence of carrageenan (Car) on the binding of AK to BSA. Results revealed that the quenching of BSA by AK involved the static quenching mechanism. The formed BSA-AK complexes were mainly maintained by hydrophobic forces and AK was located within the hydrophobic cavity of BSA. Compared to free AK or AK only complexed with BSA, a higher absorption intensity of AK was observed for the formed BSA-AK-Car complexes, indicating changes in the microenvironment of AK. This was confirmed by the increase in the α-helix content of BSA after the formation of BSA-AK-Car complexes. Hydrogen bond, van der Waals, and electrostatic interactions were verified to be the primary forces preserving the BSA-AK-Car complexes. Moreover, the antioxidant potential of Monascus-fermented products rich in AK (denoted as Mps), namely BSA-Mps and BSA-Mps-Car was evaluated. The antioxidant activity of Mps was negatively impacted by BSA, while the addition of Car could enhance the antioxidant capacity of BSA-Mps-Car complexes. Meanwhile, Mps showed a protective effect against free radical-induced oxidation damage to BSA, and Car could further improve this effect.

Iron (II) and Zinc (II) complexes of gemifloxacin mesylate: synthesis, characterization, serum binding profiling, and evaluation of antimicrobial activity

Gemifloxacin mesylate is a synthetic fluoroquinolone derived antimicrobial agent. Two metal complexes of gemifloxacin mesylate were synthesized viz. gemifloxacin-Fe and gemifloxacin-Zn. The formed complexes were characterized by using TLC, TGA and FT-IR spectra analyses. The formed complexes were then evaluated for antimicrobial activity. The newly formed two complexes were assessed against nine bacterial and one fungal strain, where gemifloxacin (30 µg/disc) was used as a reference standard. The new complexes showed better activity than the standard reference drug against Klebsiella pneumoniae among nine bacterial strains. But two bacterial species, Acinetobacter lwoffii and Enterococcus faecium, and the fungal strain Candida albicans showed complete resistance to the newly synthesized complexes and the reference standard. The drug protein interaction with the Bovine Serum Albumin (BSA) was also studied. The interaction mechanism was explored, suggesting that gemifloxacin and its Zn (II) complex interact with BSA via a static process while the Fe (II) complex interacts via a dynamic process and the lower Ksv value also indicated that drug -BSA complexes were formed in ground-state. J. Bangladesh Acad. Sci. 46(2); 203-211: December 2022

Chiral recognition of tryptophan enantiomer based on the electrode modified by polyaniline adsorption bovine serum albumin complex.

A chiral sensing platform was constructed via adsorptive functionalization of ammonium persulfate doped polyaniline (APS-DPANI) with bovine serum albumin (BSA). The novelty of this work is the construction of such chiral interface with adsorption principle. The material has been characterized by scanning electron microscope, Fourier transform infrared and X-ray photoelectron spectroscopy, and thermogravimetric and water contact angle analyses. It displayed considerable stability in multi-run cyclic voltammetric scanning. Moreover, the superior conductivity of APS-DPANI and the decent binding ability of BSA endowed this sensing platform with an excellent recognition effect for tryptophan (Trp) enantiomers in the differential pulse voltammetry (DPV) test. The recognition was highly reproducible, and the detection limits for L- and D-isomer were 0.071 and 0.0478 mM, respectively.

3-Hydroxyflavone - bovine serum albumin complex used as fluorescent probe for the detection of pepsin

In this study, a method for the determination of pepsin using a 3-hydroxyflavone (3-HF)-bovine serum albumin (BSA) complex as a fluorescent probe was established. Under acidic conditions, 3-HF can react with BSA to emit strong green fluorescence, which involves an excited state intramolecular proton transfer and fluorescence resonance energy transfer. After pepsin is added, the fluorescence of the 3-HF-BSA system is quenched due to the hydrolysis of BSA. The degree of 3-HF-BSA fluorescence quenching has a good linear relationship with the concentration of pepsin in the range of 10–80 μg/mL. The correlation coefficient was R2 = 0.9932 and the detection limit was 2.41 μg/mL. This method has high selectivity and good stability and has been used to determine pepsin in real samples with satisfactory results.

Foaming properties of the complex of chitooligosaccharides and bovine serum albumin and its application in angel cake

The complex of chitooligosaccharides (COS) and protein is worthy of investigation, as COS is an important functional ingredient and the functional properties of protein might be improved after complexation. Using bovine serum albumin (BSA) as the model protein, this work explored the foaming properties and interfacial behavior of COS-BSA complex, as well as its application as a substitute for egg whites in angel cake. The effects of pH and mixing ratio ( r ) on the foam properties and interfacial behavior of COS-BSA complex were studied using dilute solution. And the effects of the COS-BSA substitution rate on the physical properties and microbial storability of the cake were studied. Compared with BSA, the foaming capacity and foam stability of COS-BSA complexes were significantly improved in the pH range of 3.0–10.0, and both the foaming capacity and foam stability of COS-BSA complexes at pH 7.0 were in the order of r 0.5 ≥ r 0.25 ≥ r 1.0 > r 2.0 > BSA > COS. Angel cake with the COS-BSA substitution rate of 25% had the best physical properties, in which COS and BSA had a synergistic effect on the specific volume and texture of the cake. The substitute of COS-BSA complex for egg white in angel cake significantly inhibited microbial growth during storage. This work provided COS-BSA complex as a substitute for egg white in cake making, and COS-BSA complex improved the physical properties of cake and inhibited microbial growth. • Chitooligosaccharides improved foaming properties of bovine serum albumin. • Good foaming of complexes was related with their surface dilatational rheology. • Cake with complex substitution rate of 25% exhibited the best physical properties. • Complex addition in cake significantly inhibited microbial growth during storage.

Characterization of Climbazole–Bovine serum albumin interaction by experimental and in silico approaches

Interactive association of an antifungal drug, climbazole (CBZ) with the carrier protein in bovine circulation, bovine serum albumin (BSA) was explored by fluorescence and absorption spectroscopy along with in silico techniques. The fluorescence and absorption spectral alterations of the protein upon addition of CBZ affirmed the complex foration between CBZ and BSA. The inverse temperature dependence behaviour of the KSV values as well as the hyperchromic result of the protein’s absorption signals characterized CBZ-triggered quenching of BSA fluorescence as the static quenching. A weak binding affinity (Ka = 3.12–1.90-× 103 M−1) was reported towards the CBZ−BSA association process. Interpretation of thermodynamic data (entropy change = +14.68 J mol−1 K−1 and enthalpy change = –15.07 kJ mol−1) and in silico analyses anticipated that hydrophobic forces, van der Waals forces and hydrogen bonds were the key intermolecular forces in the complex stabilization. Inclusion of CBZ to BSA produced microenvironmental perturbations around Tyr and Trp residues, and also significantly defended temperature-induced destabilization of BSA. The binding locus of CBZ was detected in the proximity of Sudlow’s sites I (subdomain IIA) and II (subdomain IIIA) of BSA, exhibiting greater preference towards site II, as revealed by competitive site-marker displacement investigations and in silico analysis. The stability of the CBZ–BSA complex was further validated by the molecular dynamics simulation assessments.

Photocatalytic Pt(IV)-Coordinated Carbon Dots for Precision Tumor Therapy.

Rapid, efficient, and precise cancer therapy is highly desired. Here, this work reports solvothermally synthesized photoactivatable Pt(IV)-coordinated carbon dots (Pt-CDs) and their bovine serum albumin (BSA) complex (Pt-CDs@BSA) as a novel orange light-triggered anti-tumor therapeutic agent. The homogeneously distributed Pt(IV) in the Pt-CDs (Pt: 17.2 wt%) and their carbon cores with significant visible absorption exhibit excellent photocatalytic properties, which not only efficiently releases cytotoxic Pt(II) species but also promotes hydroxy radical generation from water under orange light. When triggered with a 589nm laser, Pt-CDs@BSA possesses the ultrastrong cancer cell killing capacities of intracellular Pt(II) species release, hydroxyl radical generation, and acidification, which induce powerful immunogenic cell death. Activation of Pt-CDs@BSA by a single treatment with a 589nm laser effectively eliminated the primary tumor and inhibited distant tumor growth and lung metastasis. This study thus presents a new concept for building photoactivatable Pt(IV)-enriched nanodrug-based CDs for precision cancer therapy.

Synthesis of a β-Arylethenesulfonyl Fluoride-Functionalized AIEgen for Activity-Based Urinary Trypsin Detection.

Trypsin is one of the most important enzymes of the digestive system produced by the pancreatic acinar cells. Abnormal trypsin activity will affect pancreatic function, resulting in the corresponding pathological changes in the human body. Herein, we present a strategy based on the ensemble of a novel dual warhead probe HPC-ESF and the natural trypsin substrate bovine serum albumin (BSA) for the detection of trypsin activity including in real urine samples. The β-arylethenesulfonyl bearing HPC-ESF is nonemissive when dissolved in aqueous solution but becomes highly fluorescent upon conjugation to BSA through covalent bond formation with nucleophilic amino acids to create the HPC-ESF:BSA sensing system. The HPC-ESF:BSA complex can be hydrolyzed in the presence of trypsin, which results in a distinct fluorescence decrease in correlation with trypsin concentration and thus allows the detection of trypsin. Compared to previous methods, our covalent approach is simple to prepare and highly reliable. Our work will provide a different avenue for researchers to design fluorescent sensors based on a covalent labeling strategy, enriching the small library of functional groups available for such applications.

Hybrid (Bovine Serum Albumin)/Poly(N-vinyl-2-pyrrolidone-co-acrylic acid)-Shelled Microbubbles as Advanced Ultrasound Contrast Agents.

Microbubbles are routinely used ultrasound contrast agents in the clinic. While a soft protein shell is commercially preferable for imaging purposes, a rigid polymer shell demonstrates prolonged agent stability. Hence, combining polymers and proteins in one shell composition can advance microbubble properties. We formulated the hybrid "protein-copolymer" microbubble shell with a complex of bovine serum albumin and an amphiphilic copolymer of N-vinyl-2-pyrrolidone and acrylic acid. The resulting microbubbles demonstrated advanced physicochemical and acoustic properties, preserving in vitro biocompatibility. Adjusting the mass ratio between protein and copolymer allowed fine tuning of the microbubble properties of concentration (by two orders, up to 1010 MBs/mL), mean size (from 0.8 to 5 μm), and shell thickness (from 28 to 50 nm). In addition, the minimum air-liquid surface tension for the "protein-copolymer" solution enabled the highest bubble concentration. At the same time, a higher copolymer amount in the bubble shell increased the bubble size and tuned duration and intensity of the contrast during an ultrasound procedure. Demonstrated results exemplify the potential of the hybrid "protein-polymer" microbubble shell, allowing tailoring of microbubble properties for image-guided applications, combining advances of each material involved in the formulation.

Conformational Changes in the BSA-LT4 Complex Induced by the Presence of Vitamins: Spectroscopic Approach and Molecular Docking.

Levothyroxine (LT4) is known for its use in various conditions including hypothyroidism. LT4 interaction with serum albumin may be influenced by the presence of vitamins. For this reason, we investigated the effect of vitamin C, vitamin B12, and folic acid on the complex of Bovine Serum Albumin with LT4 (BSA-LT4). UV-Vis spectroscopy was used to monitor the influence of vitamins on the BSA-LT4 complex. Fluorescence spectroscopy revealed a static quenching mechanism of the fluorescence of BSA-LT4 complex by the vitamin C and folic acid and a combined mechanism for vitamin B12. The interaction of vitamin C and folic acid with BSA-LT4 was moderate, while the binding of vitamin B12 was much stronger, extending the storage time of LT4 in blood plasma. Synchronous fluorescence found that the vitamins were closer to the vicinity of Trp than to Tyr and the effect was more pronounced for the binding of vitamin B12. The thermal stability of the BSA-LT4 complex was more evident, but no influence on the stability of BSA-LT4 complex was obtained for vitamin C. Molecular docking studies showed that vitamin C and folic acid bound the same site of the protein, while vitamin B12 bonded to a different site.

Bovine serum albumin with gallic acid: Molecular modeling and physicochemical profiling

Introduction. Gallic acid is a biologically active natural compound with strong antioxidant properties. Gallic acid is highly soluble and stable. It is known to increase the thermal stability of protein. However, its bioavailability is low, but interaction with proteins can solve this problem. Bovine serum albumin can bind various ligands, including polyphenols. The resulting complex of gallic acid and bovine serum albumin can become a promising functional food additive.
 Study objects and methods. This research featured in silico molecular modeling of gallic acid and bovine serum albumin using the HyperChem program. The methods of infrared spectrometry, potentiometry, and sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) made it possible to describe the physicochemical profile of the complex.
 Results and discussion. The molecular modeling confirmed that hydrophobic interactions were responsible for the chemical bond between gallic acid and bovine serum albumin. The SDS-PAGE test showed that the protein molecule remained intact. The reducing properties of the complex grew as the concentration of gallic acid increased. At 100 mg/L of gallic acid, the reducing properties were 7.8 ± 1.3 mg/L equivalent of gallic acid. At 200 and 300 mg/L, the values reached 15.90 ± 2.65 and 23.30 ± 5.05 mg/L, respectively. The IR spectrometry revealed a significant difference between the samples with different concentrations of gallic acid.
 Conclusion. The research managed to predict the properties of the complex of bovine serum albumin and gallic acid during its formation. The resulting complex had the highest reducing properties at 0.69 g of bovine serum albumin and 300 mg of gallic acid. The obtained parameters can be used in the food industry to develop new food additives.

The synthesis and application of a colour-switch β-arylethenesulfonyl fluoride fluorescent probe in the detection of serum albumin

Proteins play a pivotal role in regulating important physiological processes and serve as important biomarkers for many diseases. Herein, we present a new strategy for bovine serum albumin (BSA) detection using a novel colour-switch fluorescent probe CPV-ESF ((E)-2-(4-((Z)-1-cyano-2-(4-(diethylamino)phenyl)vinyl)phenyl)ethene-1-sulfonyl fluoride). CPV-ESF reacts with nucleophilic amino acids of BSA via 1,4-Michael addition click chemistry to create a covalently linked CPV-ESF:BSA complex, which can be easily detected by a fluorescence colour-switch response. The sensing mechanism, sensitivity and selectivity of CPV-ESF for BSA detection as well as its application for cell imaging have been investigated.

Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane.

Gene therapy is a suitable alternative to chemotherapy due to the complications of drug resistance and toxicity of drugs, and is also known to reduce the occurrence of cellular mutation through the use of gene carriers. In this study, gene carrier nanoparticles with minimal toxicity and high transfection efficiency were fabricated from a biocompatible and biodegradable polymer, l-tyrosine polyurethane (LTU), which was polymerized from presynthesized desaminotyrosyl tyrosine hexyl ester (DTH) and polyethylene glycol (PEG), by using double emulsion and solvent evaporation techniques, resulting in the formation of porous nanoparticles, and then used to evaluate their potential biological activities through molecular controlled release and transfection studies. To assess cellular uptake and transfection efficiency, two model drugs, fluorescently labeled bovine serum albumin (FITC-BSA) and plasmid DNA-linear polyethylenimine (LPEI) complex, were successfully encapsulated in nanoparticles, and their transfection properties and cytotoxicities were evaluated in LX2 as a normal cell and in HepG2 and MCF7 as cancer cells. The morphology and average diameter of the LTU nanoparticles were confirmed using light microscopy, transmission electron microscopy, and dynamic light scattering, while confocal microscopy was used to validate the cellular uptake of FITC-BSA-encapsulated LTU nanoparticles. Moreover, the successful cellular uptake of LTU nanoparticles encapsulated with pDNA-LPEI and the high transfection efficiency, confirmed by gel electrophoresis and X-gal assay transfection, indicated that LTU nanoparticles had excellent cell adsorption ability, facilitated gene encapsulation, and showed the sustained release tendency of genes through transfection experiments, with an optimal concentration ratio of pDNA and LPEI of 1:10. All the above characteristics are ideal for gene carriers designed to transport and release drugs into the cytoplasm, thus facilitating effective gene therapy.

Color-tunable, self-healing albumin-based lanthanide luminescent hydrogels fabricated by reductant-triggered gelation

Luminescent hydrogels show extensive applications in many fields because of their excellent optical properties. Although there are many matrixes used to prepare luminescent hydrogels, the synthesis of protein-based luminescent hydrogels is still urgently needed to explore due to their good biodegradability and biocompatibility. In this work, a color-tunable, self-healing protein-based luminescent hydrogel consisting of bovine serum albumin (BSA) and lanthanide complexes is prepared via reductant-triggered gelation. Firstly, a bifunctional organic ligand named 4-(phenylsulfonyl)-pyridine-2,6-dicarboxylic acid (4-PSDPA) is synthesized, which can react with thiol groups and effectively sensitize the luminescence of Eu3+ and Tb3+ ions. Then, the BSA is treated with a reducing agent tris(2-carboxyethyl)phosphine (TCEP) to produce thiol groups. And the newly formed thiol groups can re-match to form disulfide bonds between two BSA molecules or react with Ln(4-PSDPA)3 complexes, resulting in the formation of an albumin-based luminescent hydrogel. Furthermore, the self-healing, biodegradability and biocompatibility of albumin-based hydrogels have also been demonstrated. We expect that the newly developed multifunctional protein-based hydrogels will find potential applications in the fields of biomedical engineering and optical devices.

COMPARATIVE STUDY OF SPECTRAL CHARACTERISTICS OF COMPLEXES OF HOECHST 33258 AND METHYLENE BLUE WITH BOVINE SERUM ALBUMIN

The comparative study on interaction of bisbenzimidazole compound Hoechst 33258 and thiazine dye methylene blue (MB) with bovine serum albumin (BSA) was carried out by spectroscopic methods. Denaturation curves as well as absorption spectra and differential absorption spectra of protein-ligand complexes
 were obtained. Denaturation temperature of albumin complexes of BSA with Hoechst 33258 was shown to decrease with the growth of concentration ratio of ligand/protein, while for MB, vice versa, denaturation temperature increases. Changes in absorption spectra and differential absorption spectra of the complexes
 of ligands with albumin were revealed, which result from the binding of these DNA-specific ligands to protein. It is supposed that at the interaction of Hoechst 33258 with BSA some loosening of protein compact structure occurs due to the partial loss of helicity of α-structures, while for MB an increase of the protein compact structure takes place.

Novel cholesterol efflux assay using immobilized liposome-bound gel beads: Confirmation and improvement for application in clinical laboratory.

Cholesterol efflux capacity (CEC), an atheroprotective function of high-density lipoprotein, is expected to be a potential biomarker for cardiovascular disease. However, CEC has not been widely introduced for application in clinical laboratories because of the complexity of the conventional CEC assay using cells and radioactive materials. Previously, we developed a novel CEC assay using immobilized liposome-bound gel beads (ILG), which solves these issues. We aimed to confirm the validation and further improve the ILG method for application in the clinical setting. Cholesterol efflux capacity values by the ILG method assayed for shorter incubation time (4h) were compared to those assayed for 16h (our previous ILG method). To investigate a reference material that can correct the variation between ILG manufacturing lots, bovine serum albumin, human gamma-globulins, and globulin complexes were evaluated. CEC values were also estimated in plasmas obtained with different anticoagulants, serum treated with freeze-thaw cycles, and serum mixed with several interference substances. The CEC of 4- and 16-h incubation times were well correlated. Globulin complexes may be used as a reference material. Plasma can be used as the specimen. The serum and stored temperature of the specimen did not largely affect CEC. Hemoglobin and chyle did not have an effect on CEC, whereas high-bilirubin serum showed elevated CEC. The effect of bilirubin was nearly canceled by subtracting basal fluorescence intensity. Present ILG method further fulfills some requirements for application in clinical laboratory. Using this reliable simple method, evaluation for clinical significance of CEC is expected.

Effect of Tetraphenylborate on Physicochemical Properties of Bovine Serum Albumin.

The binding interactions of bovine serum albumin (BSA) with tetraphenylborate ions ([B(Ph)4]−) have been investigated by a set of experimental methods (isothermal titration calorimetry, steady-state fluorescence spectroscopy, differential scanning calorimetry and circular dichroism spectroscopy) and molecular dynamics-based computational approaches. Two sets of structurally distinctive binding sites in BSA were found under the experimental conditions (10 mM cacodylate buffer, pH 7, 298.15 K). The obtained results, supported by the competitive interactions experiments of SDS with [B(Ph)4]− for BSA, enabled us to find the potential binding sites in BSA. The first site is located in the subdomain I A of the protein and binds two [B(Ph)4]− ions (logK(ITC)1 = 7.09 ± 0.10; ΔG(ITC)1 = −9.67 ± 0.14 kcal mol−1; ΔH(ITC)1 = −3.14 ± 0.12 kcal mol−1; TΔS(ITC)1 = −6.53 kcal mol−1), whereas the second site is localized in the subdomain III A and binds five ions (logK(ITC)2 = 5.39 ± 0.06; ΔG(ITC)2 = −7.35 ± 0.09 kcal mol−1; ΔH(ITC)2 = 4.00 ± 0.14 kcal mol−1; TΔS(ITC)2 = 11.3 kcal mol−1). The formation of the {[B(Ph)4]−}–BSA complex results in an increase in the thermal stability of the alfa-helical content, correlating with the saturation of the particular BSA binding sites, thus hindering its thermal unfolding.

Tracking Zone-wise perturbation during unfolding of some globular proteins using Eu(III) complex of Tetracycline as a probe exhibiting Stark splitting

Enhanced ‘Antenna effect’ of a suitably designed ternary complex of Eu(III), Tetracycline hydrochloride (TC) and globular proteins viz bovine serum albumin (BSA), human serum albumin (HSA) and β-lactoglobulin A (BLGA) in aqueous medium is employed to characterize the different partially unfolded states along with investigation of the micro- heterogeneous environment of the proteins during their stepwise unfolding. The zone-wise perturbation for the proteins upon denaturation by Urea and Guanidine hydrochloride (Gdn. HCl) is followed by the emission of Eu(III) through ‘Antenna Effect’ and that of the tryptophan (Trp) residues of the proteins as a function of denaturants both by steady state and time resolved emission study. With Gdn. HCl as denaturant, both BSA and BLGA show quenching of Eu(III) emission compared to pure protein while HSA exhibits an enhancement of antenna effect during unfolding as compared to that in its absence. In the presence of Urea, HSA and BSA show enhancement of antenna effect accompanied by Stark splitting of the 5D0→7F2 transition of Eu(III) although BLGA follows the similar pattern of quenching of Eu(III) emission as observed with Gdn. HCl without any Stark splitting. The proteins exhibit a two state transition with ΔGD values of ~ 2–3 kcal mol−1. Thus the use of Eu(III) emission as an efficient probe is advocating here to rationalize the microenvironment of the proteins during their stepwise unfolding.

Insights into the binding interaction mechanism of 12,12-dihydrochromeno[2,3-c]isoquinolin-5-amine in bovine serum albumin and prostaglandin H2 synthase-1: A biophysical approach

The current study investigates the newly synthesized 12,12-dihydrochromeno[2,3-c]isoquinolin-5-amine (drug) molecule binding interaction mechanism with bovine serum albumin (BSA) and prostaglandin H2 synthase-1(PTGS1), using biophysical techniques. The emission spectroscopy shows a possibility of static quenching mechanism between BSA and drug complex and the thermo dynamical parameter result suggests hydrophobic interactions between BSA and drug. The excitation emission matrix and circular dichroism analysis concludes the presence of drug in BSA molecule. Molecular docking studies for the complex of drug shows good binding affinity in BSA, PTGS1 complex system and the BSA-drug computational docking score is almost in the same order of experimentally determined values. The molecular dynamics simulations were performed to understand drug binding stability in BSA, PTGS1 complex system for 50 ns. The density functional theory calculations were also performed to understand the intermolecular interaction mechanism of free drug and drug in BSA, PTGS1 binding active site environment.