Entropy (ΔS), enthalpy (ΔH) and heat capacity (ΔCp) changes attending the oxytocin interaction with its two binding sites on myometrial cell membranes in sheep were derived from the temperature dependence of Kd values. The high affinity oxytocin site (Kd on the order of 10(-9)mol l(-1), 25 °C), ascribed to the oxytocin receptor (OXTR), is entropy-driven in the temperature range 0-37 °C. Enthalpy component prevails as a driving force in the binding to the low affinity site (Kd ≈ 10(-7)) within the higher temperature range. ΔCp values in both cases do not differ significantly from zero but become highly relevant in the presence of a GTP analog (10(-4)M GTP-γS). Under these conditions, ΔCp in the low site interaction becomes negative and ΔS is shifted toward negative values (enthalpy drift); ΔCp of the high affinity site rises to a high positive value and the interaction is even more strongly entropy driven. Atosiban, a competitive antagonist of oxytocin at OXTR displays a single significant binding site on myometrial cells (Kd about 10(-7)mol l(-1)). Thermodynamic profiles of atosiban and the low affinity oxytocin site show conspicuous similarities, indicating that the inhibitor is bound to the low affinity site, and not, with a lower affinity, to the putative receptor protein. It is suggested that the interaction of oxytocin with its responding system on myometrial membranes follows in two distinct steps that are likely to be associated with several independent binding domains in the GPCR receptor.

• MeSH
• Arginine Vasopressin metabolism   MeSH
• Cell Membrane metabolism   MeSH
• Myometrium metabolism   MeSH
• Sheep MeSH
• Oxytocin metabolism   MeSH
• Receptors, Oxytocin metabolism   MeSH
• Temperature MeSH
• Thermodynamics MeSH
• Vasotocin analogs & derivatives   metabolism   MeSH
• Binding Sites MeSH
• Animals MeSH
• Check Tag
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

The binding ability of a drug to plasma proteins influences the pharmacokinetics of a drug. As a result, it is a very important issue in new drug development. In this study, affinity capillary electrophoresis, capillary electrophoresis with frontal analysis, and Hummel Dreyer methods with internal and external calibration were used to study the affinity between bovine serum albumin and salicylic acid. The binding constant was measured by all these approaches including the equilibrium dialysis, which is considered to be a reference method. The comparison of results and other considerations showed the best electrophoretic approach to be capillary electrophoresis-frontal analysis, which is characterized by the high sample throughput with the possibility of automation, very small quantities of biomacromolecules, simplicity, and a short analysis time. The mechanism of complex formation was then examined by capillary electrophoresis with frontal analysis. The binding parameters were determined and the corresponding thermodynamic parameters such as Gibbs free energy ΔG(0), enthalpy ΔH(0), and entropy changes ΔS(0) at various temperatures were calculated. The results showed that the binding of bovine serum albumin and salicylic acid was spontaneous, and that hydrogen bonding and van der Waals forces played a major role in the formation of the complex.

• MeSH
• Automation * MeSH
• Electrophoresis, Capillary methods   MeSH
• Calibration MeSH
• Pharmaceutical Preparations metabolism   MeSH
• Proteins metabolism   MeSH
• Thermodynamics MeSH
• Protein Binding MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

Combined effects of temperature and mobile phase on the reversed phase chromatographic behavior of alkylbenzenes and simple substituted benzenes were investigated on a Blaze C8 polydentate silica-based column, showing improved resistance against hydrolytic breakdown at temperatures higher than 60 degrees C, in comparison to silica-based stationary phases with single attachment sites. For better insight into the retention mechanism on polydentate columns, we determined the enthalpy and entropy of the transfer of the test compounds from the mobile to the stationary phase. The enthalpic contribution dominated the retention at 80% or lower concentrations of methanol in the mobile phase. Entropic effects are more significant in 90% methanol and in acetonitrile-water mobile phases. Anomalies in the effects of mobile phase on the enthalpy of retention of benzene, methylbenzene and polar benzene derivatives were observed, in comparison to regular change in enthalpy and entropy of adsorption with changing concentration of organic solvent and the alkyl length for higher alkylbenzenes. The temperature and the mobile phase effects on the retention are practically independent of each other and--to first approximation--can be described by a simple model equation, which can be used for optimization of separation conditions.

• MeSH
• Acetonitriles chemistry   MeSH
• Benzene Derivatives chemistry   MeSH
• Models, Chemical MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Linear Models MeSH
• Methanol chemistry   MeSH
• Reproducibility of Results MeSH
• Temperature MeSH
• Thermodynamics MeSH
• Chromatography, High Pressure Liquid methods   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

Hydrophilic divinylbenzene (DVB) (Bakerbond) has surfaced as a promising sorbent for active sampling of analytes from aqueous matrices over a very broad polarity range. Given this, hydrophilic DVB may likewise offer potential for passive sampling, if sorbent/water partitioning coefficients (Ksw) were to be available. In this work, static exposure batch experiments were performed to quantitatively study the equilibrium sorption of 131 environmentally relevant organic contaminants (P values ranging from -1.30 to 9.85) on hydrophilic DVB. The superior affinity of hydrophilic DVB, as compared to Oasis HLB, for compounds with a broad polarity range was confirmed by functional Fourier-transform infrared spectroscopy and Raman characterization, demonstrating the presence of carboxyl moieties. Concentration effects were studied by increasing compound concentrations in mixture experiments and resulted in the steroidal endocrine disrupting compounds in higher Ksw, while lower Ksw were obtained for the (alkyl)phenols, personal care products, pesticides, pharmaceuticals, and phthalates. Nevertheless, Ksw remained constant in the said design for equilibrium water concentrations at environmentally relevant seawater levels. An independent analysis of thermodynamic parameters (change in enthalpy, entropy, and Gibbs free energy) revealed the nature of the main partitioning processes. While polar (log P < 4) compounds were mainly served by physisorption, nonpolar (log P > 4) compounds also exhibited binding by multiple hydrogen bonding. In conclusion, this research facilitates the future application of hydrophilic DVB for active as well as passive sampling in the analysis of organic contaminants for monitoring purposes and for toxicity testing.

• MeSH
• Water Pollutants, Chemical * MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Pesticides * MeSH
• Vinyl Compounds MeSH
• Publication type
• Journal Article MeSH

A semiempirical quantum mechanical PM6-DH2 method accurately covering the dispersion interaction and H-bonding was used to score fifteen structurally diverse CDK2 inhibitors. The geometries of all the complexes were taken from the X-ray structures and were reoptimised by the PM6-DH2 method in continuum water. The total scoring function was constructed as an estimate of the binding free energy, i.e., as a sum of the interaction enthalpy, interaction entropy and the corrections for the inhibitor desolvation and deformation energies. The applied scoring function contains a clear thermodynamical terms and does not involve any adjustable empirical parameter. The best correlations with the experimental inhibition constants (ln K (i)) were found for bare interaction enthalpy (r (2) = 0.87) and interaction enthalpy corrected for ligand desolvation and deformation energies (r (2) = 0.77); when the entropic term was considered, however, the correlation becomes worse but still acceptable (r (2) = 0.52). The resulting correlation based on the PM6-DH2 scoring function is better than previously published function based on various docking/scoring, SAR studies or advanced QM/MM approach, however, the robustness is limited by number of available experimental data used in the correlation. Since a very similar correlation between the experimental and theoretical results was found also for a different system of the HIV-1 protease, the suggested scoring function based on the PM6-DH2 method seems to be applicable in drug design, even if diverse protein-ligand complexes have to be ranked.

• MeSH
• Cyclin-Dependent Kinase 2 antagonists & inhibitors   metabolism   MeSH
• Protein Kinase Inhibitors chemistry   pharmacology   MeSH
• Quantum Theory MeSH
• Humans MeSH
• Ligands MeSH
• Models, Molecular MeSH
• Drug Design MeSH
• Thermodynamics MeSH
• Protein Binding MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Biophysics MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• Učební osnovy. Vyučovací předměty. Učebnice
• NML Fields
• fyzika, biofyzika
• NML Publication type
• učebnice vysokých škol

• MeSH
• Health Physics MeSH
• Biophysics MeSH
• Electrophysiology MeSH
• Molecular Structure MeSH
• Spectrum Analysis MeSH
• Thermodynamics MeSH
• Publication type
• Textbook MeSH

• Conspectus
• Biochemie. Molekulární biologie. Biofyzika
• NML Fields
• fyzika, biofyzika

• Keywords
• Lékařské obory,
• MeSH
• Biophysics MeSH

• NML Fields
• fyzika, biofyzika

• MeSH
• Biophysics MeSH

• Conspectus
• Lékařské vědy. Lékařství
• Učební osnovy. Vyučovací předměty. Učebnice
• NML Fields
• fyzika, biofyzika
• NML Publication type
• učebnice vysokých škol

UNLABELLED: We report enzymologic, thermodynamic and structural analyses of a series of six clinically derived mutant HIV proteases (PR) resistant to darunavir. As many as 20 mutations in the resistant PRs decreased the binding affinity of darunavir by up to 13 000-fold, mostly because of a less favorable enthalpy of binding that was only partially compensated by the entropic contribution. X-ray structure analysis suggested that the drop in enthalpy of darunavir binding to resistant PR species was mostly the result of a decrease in the number of hydrogen bonds and a loosening of the fit between the inhibitor and the mutated enzymes. The favorable entropic contribution to darunavir binding to mutated PR variants correlated with a larger burial of the nonpolar solvent-accessible surface area upon inhibitor binding. We show that even very dramatic changes in the PR sequence leading to the loss of hydrogen bonds with the inhibitor could be partially compensated by the entropy contribution as a result of the burial of the larger nonpolar surface area of the mutated HIV PRs. DATABASE: Atomic coordinates and structure factors for the crystal structures PRwt-DRV and PRDRV2-DRV complex have been deposited in the Protein Data Bank under accession codes 4LL3 and 3TTP, respectively. STRUCTURED DIGITAL ABSTRACT: • PR and PR bind by x-ray crystallography (View interaction).

• MeSH
• HIV Protease chemistry   genetics   metabolism   MeSH
• HIV Protease Inhibitors chemistry   pharmacology   MeSH
• Molecular Sequence Data MeSH
• Mutation * MeSH
• Amino Acid Sequence MeSH
• Molecular Docking Simulation * MeSH
• Sulfonamides chemistry   pharmacology   MeSH
• Thermodynamics MeSH
• Protein Binding MeSH
• Drug Resistance, Viral genetics   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH