Binding Sites Of Serum Albumin Research Articles

A cobalt coordination complex binds on a unique binding site between domain-I and domain-III of serum albumin

Serum albumin binding influences physiological effects and pharmacokinetics of drugs. Major binding sites in serum albumin are sudlow sites I and II, located on domains IIA and IIIA, respectively. However, groove between domain I and III as a binding site is not well studied. In this work, we studied the binding of a cobalt coordination complex (DCoP) with bovine serum albumin (BSA) using multi-spectroscopic methods and molecular docking. DCoP strongly interacts with BSA with association constant of ∼7 × 105 M−1. The binding was found to significantly alter protein's conformation and unfolding pathway. Results of this study shows that DCoP binds at the groove formed between domains III and I. We conclude that the groove is a binding site for large molecules and coordination complexes. The knowledge of unique binding site of serum albumin enlightens the comprehensive role of protein as a carrier biomacromolecule.

Coordination compounds of cobalt(II) with carboxylate non-steroidal anti-inflammatory drugs: structure and biological profile.

Fourteen cobalt(II) complexes with the non-steroidal anti-inflammatory drugs sodium meclofenamate, tolfenamic acid, mefenamic acid, naproxen, sodium diclofenac, and diflunisal were prepared in the presence or absence of a series of nitrogen-donors (namely imidazole, pyridine, 3-aminopyridine, neocuproine, 2,2'-bipyridine, 1,10-phenanthroline and 2,2'-bipyridylamine) as co-ligands and were characterised by spectroscopic and physicochemical techniques. Single-crystal X-ray crystallography was employed to determine the crystal structure of eight complexes. The biological profile of the complexes was investigated regarding their interaction with serum albumins and DNA, and their antioxidant potency. The interaction of the compounds with calf-thymus DNA takes place via intercalation. The ability of the complexes to cleave pBR322 plasmid DNA at the concentration of 500 μM is rather low. The complexes demonstrated tight and reversible binding to human and bovine serum albumins and the binding site of bovine serum albumin was also examined. In order to assess the antioxidant activity of the compounds, the in vitro scavenging activity towards free radicals, namely 1,1-diphenyl-picrylhydrazyl and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), and their ability to reduce H2O2 were studied.

Binuclear nitrosyl iron complex with 4-acetamidothiophenolyl: Synthesis and study of its decomposition in a system with glutathione and albumin

In this work, we synthesized a new binuclear nitrosyl iron complex of the family of Roussin's red salt ester [Fe2(C8H8NOS)2(NO)4] (complex 1) with 4-acetamidothiophenol (a structural analog of acetaminophen), which generates NO during hydrolysis and is promising for treatment socially significant diseases. It has been established that solvents have a significant effect on the process of its decomposition: in a buffer solution, complex 1 retains its binuclear structure, while in DMSO it forms mononuclear iron-nitrosyl products. Reduced glutathione does not replace 4-acetamidothiophenol ligands in the structure of complex 1. In addition, complex 1 and its products are not coordinated with the known binding sites of bovine serum albumin, including Cys34. It was found that the studied complex 1 is adsorbed on the protein surface due to weak intermolecular interactions, which leads to a decrease in the rate of decomposition, and, as a result, to a longer generation of NO.Thus, it has been shown for the first time that compound 1 and its decay products do not form coordination complexes with the biosubstrates studied in this work. It can be assumed that glutathione will not participate in the transformation of complex 1 in vivo, while albumin, due to the effective stabilization of the complex on the surface, can act as its carrier to therapeutic targets.

Multi-cavity molecular descriptor interconnections: Enhanced protocol for prediction of serum albumin drug binding

The role of human serum albumin (HSA) in the transport of molecules predicates its involvement in the determination of drug distribution and metabolism. Optimization of ADME properties are analogous to HSA binding thus this is imperative to the drug discovery process. Currently, various in silico predictive tools exist to complement the drug discovery process, however, the prediction of possible ligand-binding sites on HSA has posed several challenges. Herein, we present a strong and deeper-than-surface case for the prediction of HSA-ligand binding sites using multi-cavity molecular descriptors by exploiting all experimentally available and crystallized HSA-bound drugs. Unlike previously proposed models found in literature, we established an in-depth correlation between the physicochemical properties of available crystallized HSA-bound drugs and different HSA binding site characteristics to precisely predict the binding sites of investigational molecules. Molecular descriptors such as the number of hydrogen bond donors (nHD), number of heteroatoms (nHet), topological polar surface area (TPSA), molecular weight (MW), and distribution coefficient (LogD) were correlated against HSA binding site characteristics, including hydrophobicity, hydrophilicity, enclosure, exposure, contact, site volume, and donor/acceptor ratio. Molecular descriptors nHD, TPSA, LogD, nHet, and MW were found to possess the most inherent capacities providing baseline information for the prediction of serum albumin binding site. We believe that these associations may form the bedrock for establishing a solid correlation between the physicochemical properties and Albumin binding site architecture. Information presented in this report would serve as critical in provisions of rational drug designing as well as drug delivery, bioavailability, and pharmacokinetics.

New anthracene-based Oxime-Palladium complexes loaded on albumin nanoparticles, in vitro cytotoxicity, mathematical release mechanism studies and biological macromolecules interaction investigation

In this research work, two new palladium complexes [trans-Pd(C15H10NOCH3)2]Cl2 (1) and [cis- Pd(C15H10NOCH3)(PPh3)2Cl]Cl (2) were synthesized using an alkoxyme ligand named isophethalaldoxime. Then structure characterization has been done by FT-IR and different NMR (1H, 13C and 31P) spectroscopy. Then, their interactions with biological macromolecules including deoxyribonucleic acid and bovine serum albumin were studied using various spectroscopic methods such as UV–Vis absorption, fluorescence emission spectroscopy and circular dichroism. The results showed the binding of the prepared complexes to the deoxyribonucleic acid via grooves and different binding sites of bovine serum albumin. Fluorescence emission data showed that the mechanism of extinction of albumin emission by these compounds is static. Competitive titration was performed on albumin with eosin-Y, ibuprofen and digoxin as site markers I, II and III. The antitumor activity and toxicity of these compounds were evaluated on cancer cell lines A549 (leukemia) and K562 by in-vitro cytotoxicity test. The IC50 values showed the good activity of these complexes in inhibiting cancer cells. In the last section, the release mechanism of synthesized complexes from albumin nanoparticles (BNPs) was investigated and theoretical calculations were performed that showed Korsmeyer-Peppas mechanism for complex (1) and Quadratic mechanism for complex (2).

Computational study to find the goat serum albumin (GSA) binding site as A-esterase

A-esterases are a classical term applied to enzymatic activity of the proteins by a mechanism not involving intermediate covalent phosphorylation, but requiring a divalent cation cofactor. Recently, a copper-dependent A-esterase activity has been identified in goat serum albumin (GSA) on the organophosphorus insecticide trichloronate. This hydrolysis was identified ex vivo with spectrophotometry and chromatography techniques. Albumin mechanism of action and catalytic site as Cu2+-dependent A-esterase are still unknown. Therefore, to know the copper bind to albumin is relevant. N-terminal sequence has been reported as the high affinity site for this cation, due to the histidine in position 3. The aim of this work in silico is to explore how occurs this metallic binding and active the esterase catalytic function. The GSA crystallized structure (PDB: 5ORI) was chosen for molecular docking and dynamics. A site-directed docking, for N-terminal site and a blind docking was done with trichloronate as ligand. Root-mean-square deviation and frequency plot was calculated to find the most frequent predicted structure and visualize the amino acids involved in binding site. The affinity energy in the blind docking (−5.80 kcal/mol) is almost twice lower than site-directed docking (−3.81 kcal/mol) and N-terminal amino acids do not appear in the most repeated structure binding site, suggesting that the protein has a site with higher affinity to the trichloronate ligand. His145 could be involved in the binding site as has been reported in previous studies.

GAS CHROMATOGRAPHY-MASS SPECTROMETRY PROFILING, ANTI-INFLAMMATORY EVALUATION, AND MOLECULAR DOCKING STUDIES OF ESSENTIAL OIL AND SCENTED EXTRACT FROM THE Porana volubilis Burm. f. FLOWER

Volatile components of Porana volubilis Burm. f. flowers from Bangkok were extracted by using steam distillation and solvent extraction prior to analysis by gas chromatography-mass spectrometry (GC-MS) and their antiinflammatory activity was assessed in vitro and in silico. Forty-two compounds were identified with essential oil. The key volatile components were detected in the hexane-scented extract. Twenty-one compounds were detected with major volatile components of octacosanaol, dodecanoic acid, nonacosane, n-hexadecanoic acid, and tetratriacontane for ethanolic scented extract. The essential oil and extract were evaluated for their in vitro anti-inflammatory effect by protein denaturation assay. The essential oil showed the most potent anti-denaturation activity with IC50 values of 537.6 g/mL. The primary components of essential oil were submitted ta molecular docking research based on their anti-inflammatory effect. The obtained results revealed that the main components were found as the crucial interactions for binding in the bovine serum albumin (BSA) binding site. The results obtained demonstrated the essential oil of P. volubilis is the potential source for the anti-inflammatory agent.

4-Dimethylamino-beta-nitrostyrene, a fluorescent solvatochromic probe to estimate the apparent dielectric constant in serum albumin: Experimental and molecular dynamics studies

The spectroscopic properties of 4-dimethylamino-beta-nitrostyrene (DANS) were theoretically and experimentally correlated with the polarity of organic solvents and applied to estimate the overall apparent dielectric constant in the binding sites of bovine serum albumin (BSA). The linear correlation between McRae's solvent polarity function and the wavelength of maximum emission (R2 = 0.8899) showed the suitability of DANS as a fluorescent solvatochromic probe. The interaction between BSA and DANS was evaluated (Ka = 3.1 × 104 mol−1 L), and based on the DANS/BSA emission, the apparent dielectric constant of BSA was estimated (ε = 5.5 ± 0.8). The experimental and theoretical absorption and emission maxima were correlated using density-functional theory. Molecular dynamics was used to calculate the mean electrostatic energy between DANS and the organic solvent molecules. These results were associated with the experimental fluorescence and applied to the BSA/DANS system, resulting in electrostatic energy (−37 kJ mol−1). The application of this model generated a dielectric constant of 5.8 ± 1.3 to BSA, which was consistent with the values obtained by McRae's solvent polarity function. The mean electrostatic energy of DANS in BSA binding sites was calculated directly from molecular dynamics simulations and resulted in −40 ± 2, −47 ± 3, and −48 ± 3 kJ mol−1 for sites I, II, and III. These results showed that the environment offered by the site I presented the value of electrostatic energy closest to the obtained with the solvatochromic model. In conclusion, DANS can be used to estimate the apparent polarity of albumins.

Binding behavior of ibuprofen-based ionic liquids with bovine serum albumin: Thermodynamic and molecular modeling studies

• Nine ibuprofen-based ionic liquids containing cations amino acid alkyl esters [AAOR] were synthesized and characterized. • The [AAOR][Ibu] are not toxic against mouse fibroblasts and keratinocytes. • Ibu-ILs bind spontaneously to bovine serum albumin with moderate affinities (Ka 1–3.7 × 10 5 L/mol) at 25 °C. • Depending on their structures, [AAOR] showed differences at the preferred binding site of bovine serum albumin. Ionic liquids based on active-pharmaceutical ingredients are an attractive alternative to classical drugs because they are able to overcome problems with drug solubility and polymorphism, hence bioavailability and as well as to propose different routes of administration and/or improve the pharmacokinetics of the drug. In this manuscript, we report the synthesis of nine amino acid alkyl esters of ibuprofen [AAOR][Ibu]. The estimated half-maximum inhibitory constants toward murine embryotic fibroblasts and human dermal keratinocytes are in the millimolar range, which makes them practically not toxic. The conversion of ibuprofen into ionic liquids does not affect the binding affinity of ibuprofen to bovine serum albumin (BSA). Molecular modeling studies have shown that bulky cations such as L-PheOEt and L-PheOPr are able to bind effectively in three different BSA binding sites. On the other hand, smaller cations preferably bind to Sudhow’s site I, the same that binds ibuprofen.

The aqueous stability and interactions of organoruthenium compounds with serum proteins, cell culture medium, and human serum.

Metal complexes bind to a wide variety of biomolecules and the control of the reactivity is essential when designing anticancer metallodrugs with a specific mode of action in mind. In this study, we used the highly cytotoxic compound [RuII(cym)(8-HQ)Cl] (cym = η6-p-cymene, 8-HQ = 8-hydroxyquinoline), the more inert derivative [RuII(cym)(8-HQ)(PTA)](SO3CF3) (PTA = 1,3,5-triaza-7-phosphaadamantane), and [RuII(cym)(PCA)Cl]Cl (PCA = pyridinecarbothioamide) as a complex with a different coordination environment about the Ru center and investigated their stability, interactions with proteins, and behavior in medium (αMEM) and human serum by capillary zone electrophoresis. The developed method was found to be robust and provides a quick and low-cost technique to monitor the interactions of such complexes with biomolecules. Each complex was found to behave very differently, emphasizing the importance of the choice of ligands and demonstrating the applicability of the developed method. Additionally, the human serum albumin binding site preference of [RuII(cym)(8-HQ)Cl] was investigated through displacement studies, revealing that the compound was able to bind to both sites I and site II, and the type of adducts formed with transferrin was determined by mass spectrometry.

Near-Infrared Electrogenerated Chemiluminescence from Simple Copper Nanoclusters for Sensitive Alpha-Fetoprotein Sensing.

Exploiting low cost, water-soluble, and near-infrared (NIR) emissive electrochemiluminophores (ECLphores) is significantly important for biological applications. In this study, bright and NIR electrogenerated chemiluminescence (ECL) emissive copper nanoclusters (Cu NCs) were synthesized through a facile one-pot wet chemical reduction method. ECL properties of obtained Cu NCs were examined in the presence of potassium persulfate, resulting in maximum intensity at 735 nm, at least 135 nm red-shifted with respect to all other Cu NCs. Electrochemistry, photoluminescence (PL), and spooling ECL spectroscopies were used to track NIR ECL emission of Cu NCs ascribed to the monomeric excited states. Due to the abundant binding sites of bovine serum albumin (BSA) to anchor target biomolecules, a sandwich-type ECL immunosensor was thus fabricated using such BSA-templated Cu NCs as tags and alpha fetoprotein antigen (AFP) as a model protein for the first time. Without assisting any signal amplification strategies, the proposed NIR ECL biosensor exhibited a wide linear range (1-400 ng mL-1) and low detection limit (0.02 ng mL-1) as well as superior selectivity and reproducibility and was successfully applied in real human serum sample determination. This work sets the stage for the development of novel non-noble metal nanoclusters for large-scale and emerging nanotechnology applications.

Photochemical Reaction of Ketoprofen with Proteinogenic Amino Acids.

Ketoprofen (KP) is one of the most popular nonsteroidal anti-inflammatory drugs; however, drug-induced photosensitivity of KP has been reported as a serious adverse effect. KP incorporated into a protein can produce an allergen under UV irradiation, which causes drug-induced photosensitivity. The photochemistry of KP with 20 kinds of proteinogenic amino acids in phosphate buffer solutions at pH 7.4 was studied by transient absorption spectroscopy. The KP carboxylate anion (KP-) gave rise to a carbanion via a decarboxylation within a laser pulse, and the carbanion yielded 3-ethylbenzophenone ketyl biradical (3-EBPH) through a proton transfer reaction. Twelve kinds of proteinogenic amino acids obviously accelerated the reaction. Structural information on the complexes of KP docked in the binding sites of human serum albumin (HSA) was obtained by molecular mechanics (MM) and molecular dynamics (MD) calculations. The photochemical reaction of KP- with amino acid residues in HSA was discussed on the basis of the experimental and calculational results. The information on the reactivity of KP with the amino acids and the stable structures of the KP-HSA complexes should be essential for understanding of the initial step for drug-induced photosensitivity.

Probing the serum albumin binding site of fenamates and photochemical protein labeling with a fluorescent dye.

Human serum albumin (HSA) can bind with numerous drugs, leading to a significant influence on drug pharmacokinetics as well as undesirable drug-drug interactions due to competitive binding. Probing the HSA drug binding site thus offers great opportunities to reveal drug-HSA binding profiles. In the present study, a fluorescent probe (E)-4-(2-(5-(4-(diphenylamino)phenyl)thiophen-2-yl)vinyl)-1-propylpyridin-1-ium (TTPy) has been prepared, which exhibits enhancement of deep-red to near-infrared (NIR) fluorescence upon HSA binding. The competitive binding assay indicated that TTPy can target the HSA binding site of fenamates, a group of non-steroidal anti-inflammatory drugs (NSAIDs), with moderate binding affinity (1.95 × 106 M-1 at 303 K). More interestingly, TTPy enables fluorescent labeling of HSA upon visible light irradiation. This study provides promising ways for HSA drug binding site identification and photochemical protein labeling.

Influence of mutations caused by radiation exposure on the bilirubin binding sites of human serum albumin

In this article we analyze the bilirubin binding sites of human serum albumin from the point of view of the secondary structure instability, as well as the effect of amino acid substitutions caused by radiation exposure on the ability of albumin to bind bilirubin-IX-alpha. Based on calculations of binding energy and inhibition constants of bilirubin-albumin complexes before and after the amino acid substitutions, it was found that amino acid substitutions have different effects on the ability of human serum albumin to bind bilirubin. Amino acid substitutions Asp269-Gly269 (Nagasaki-1), Glu354-Lys354 (Hiroshima-1), Asp375-Asn375 (Nagasaki-2) reduce the binding free energy of bilirubin with human serum albumin, and the amino acid substitutions His3-Gln3 (Nagasaki-3) and Glu382-Lys382 (Hiroshima-2) increase it during molecular docking with the corresponding areas of the protein surface. The inhibition constants are significantly higher than with known binding sites. In general, mutations caused by radiation exposure cannot effect on bilirubin binding sites of human serum albumin, since the amino acid residues that are replaced do not interact with the amino acid residues from the binding sites (Leu115, Arg117, Phe134, Tyr138, Ile142, Phe149, Phe157, Tyr161, Arg186, Lys190, Lys240, Arg222). All amino acid residues from known binding sites are located in stable elements of the secondary structure of human serum albumin.The data obtained are important for understanding the impact of radiation exposure on the development of bilirubin encephalopathy in the population of the Chernobyl region and Japan.

Elucidating the Molecular Interactions between Uremic Toxins and the Sudlow II Binding Site of Human Serum Albumin.

Protein bound uremic toxins (PBUTs) have been correlated to poor clinical outcomes for patients with chronic kidney disease (CKD) and are not susceptible to the traditional dialysis techniques. Several PBUTs are known to bind strongly with the primary drug carrying sites of human serum albumin (HSA): Sudlow site I and Sudlow site II. A detailed energetic and structural description of PBUT interactions with these binding sites would provide useful insight into the design of materials that specifically displace and capture PBUTs. In this work, we used molecular dynamics (MD) simulations to study in atomistic detail four PBUTs bound in Sudlow site II. Specifically, we used the experimentally resolved X-ray structure of simulated indoxyl sulfate (IS) bound to Sudlow site II (PDB ID: 2BXH) to generate initial binding poses for p-cresyl sulfate (pCS), indole-3-acetic acid (IAA), and hippuric acid (HA). We calculated the interaction energy between toxin and protein in MD simulations and performed mean shift clustering on the collection of molecular structures from MD to identify the primary binding modes of each toxin. We find that all four toxins are primarily stabilized by electrostatic interactions between their anionic moiety and the hydrophilic residues in Sudlow site II. We observed transience in the strongest toxin-protein interaction, a charge-pairing with the positively charged R410 residue. We confirm the finding that the primary binding pose of IS in Sudlow site II is stabilized by a hydrogen bond with the carbonyl oxygen of L430 and find that this is also true for IAA. We provide insight into the chemical functional groups that might be incorporated to improve the specificity of synthetic materials for PBUT capture. This work represents a next step toward the de novo design of solutions to the problem of PBUT management in CKD patients.

Unveiling the interaction profile of rosmarinic acid and its bioactive substructures with serum albumin

Rosmarinic acid, a phytochemical compound, bears diverse pharmaceutical profile. It is composed by two building blocks: caffeic acid and a salvianic acid unit. The interaction profile, responsible for the delivery of rosmarinic acid and its two substructure components by serum albumin remains unexplored. To unveil this, we established a novel low-cost and efficient method to produce salvianic acid from the parent compound. To probe the interaction profile of rosmarinic acid and its two substructure constituents with the different serum albumin binding sites we utilised fluorescence spectroscopy and competitive saturation transfer difference NMR experiments. These studies were complemented with transfer NOESY NMR experiments. The thermodynamics of the binding profile of rosmarinic acid and its substructures were addressed using isothermal titration calorimetry. In silico docking studies, driven by the experimental data, have been used to deliver further atomic details on the binding mode of rosmarinic acid and its structural components.

The displacement study of 99m Tc-DTPA-Human serum albumin binding in presence of furosemide and metformin by using equilibrium dialysis and molecular docking.

The 99m Tc-DTPA (Technetium99m diethylenetriaminepentaacetic acid), is a radiopharmaceutical used in renal scintigraphy. The human serum albumin (HSA) binding site(s) for the 99m Tc-DTPA have never been characterized. This study will cover in vitro the binding rates of 99m Tc-DTPA on HSA and the 99m Tc-DTPA competition interactions with two drugs having known human serum albumin binding sites. Furosemide (FUR) and metformin (MET) were added to 99m Tc-DTPA solution (weight ratios 1/1 vol:vol) followed by the quantification of 99m Tc-DTPA binding rates to HSA (40 g/L) using equilibrium dialysis and the qualification of this binding using Molecular Modeling methods. The 99m Tc-DTPA binding rates to human serum albumin increased with the highest concentration. Both drugs FUR and MET displaced 99m Tc-DTPA binding. 99m Tc-DTPA could bind to human serum albumin in many locations in site I and I-II, but strongly bound to site I through hydrogen bonds.

Influence of pH and ionic strengths on bilirubin binding to sheep serum albumin

Despite great degree of similarity in the structure and conformation of different serum albumins, they often show marked difference in the ligand binding properties. These differences and similarities would be useful in understanding the molecular basis of ligand binding to serum albumins. In the present communication binding of bilirubin with sheep and bovine serum albumin have been studied by visible difference spectroscopy as a function of pH and ionic strength. The results suggest that the structure of bilirubin binding site in sheep serum albumin is similar to that in bovine serum albumin and that complex between sheep serum albumin and bilirubin is stabilized by electrostatic interactions. Keywords: Sheep serum albumin, Bovine serum albumin, Bilirubin binding, ionic strength, pH.

Spectroscopic overview of quercetin and its Cu(II) complex interaction with serum albumins.

Introduction: Flavonoids are widely used as dietary supplements, and thus, play a significant role in the research field. In recent time, the interaction of flavonoid-metal complexes with serum albumin (SA) has widely been studied since the complexation poses a significant impact on biological activities. Additionally, the binding nature of flavonoids with SA gets modified in the presence of metal ions. Methods: In the present review, we studied the interaction of quercetin (Qu), a well-known flavonoid, and its Cu2+ complexes with SA to provide sufficient information about the beneficial role of metal-flavonoid complexes over free flavonoids. Results: Complexation with Cu(II) ion may alter the mode of binding of Qu with SAs. The strength of binding might be increased in the presence of Cu(II) as evidenced by the binding constant calculation. However, the drug binding site in bovine serum albumin (BSA) and human serum albumin (HSA) are not altered during the complexation process. Conclusion: To enhance the pharmaceutical outcomes of Qu molecules, one may use Qu-Cu(II) complex for the development and delivery of the small molecules.

Novel antimycobacterial C-21 amide derivatives of the antibiotic fusidic acid: synthesis, pharmacological evaluation and rationalization of media-dependent activity using molecular docking studies in the binding site of human serum albumin.

Fusidic acid is a natural product antibiotic used clinically, primarily against staphylococcal infections. It has also exhibited antimycobacterial activity against Mycobacterium species, including Mycobacterium tuberculosis (Mtb). Novel C-21 fusidic acid amides were synthesized and evaluated for antimycobacterial activity in a drug repositioning approach for tuberculosis. The synthesized compounds exhibited good potency in MB7H9/CAS medium albeit showing low to no activity in MB7H9/ADC medium. The fusidic acid ethanamides were, generally, the most potent of the analogues evaluated for antimycobacterial activity (MIC90 < 10 μM) in the MB7H9/CAS medium. The lack of activity in the MB7H9/ADC medium was supported by strong binding interactions in the fusidic acid binding site of the human serum albumin (HSA) protein. The most potent antimycobacterial analogue was the N-(4-sulfamoylbenzyl)fusidic acid amide (1.26) with an MIC90 value of 2.71 μM.