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Binding Parameters (binding + parameter)
Selected AbstractsBinding of olive oil phenolics to food proteinsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2005Are Hugo Pripp Abstract In this paper we investigate the interaction of phenolics extracted from olive oil with different food proteins (sodium caseinate, bovine serum albumin, ,-lactoglobulin and gelatin). Binding parameters are estimated using different experimental techniques: gel filtration, HPLC, isothermal titration calorimetry and NMR diffusion measurements. For comparison, the binding properties of gallic acid and tannic acid are also studied. The affinity of olive oil phenolics for the different food proteins is found to be relatively weak (compared with tannic acid). Binding constants are measured for the different phenolics in the extract: tyrosol and hydroxytyrosol do not (or very weakly) bind to the proteins, whereas other phenolics in the extract had binding constants of the order 102,104M,1. The binding parameters determined have been discussed in relation to the possible effect of proteins on sensory properties (bitterness) of food emulsions containing olive oil. Copyright © 2004 Society of Chemical Industry [source] Effects of altered plasma ,-1-acid glycoprotein levels on pharmacokinetics of some basic antibiotics in pigs: simulation analysisJOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 6 2001M. Kuroha Effects of altered plasma , -1-acid glycoprotein (AGP) levels on pharmacokinetic parameters of basic antimicrobials, erythromycin (EM), lincomycin (LM) and clindamycin (CM) were evaluated in pigs by simulation analysis. Intravenous (i.v.) injections of EM, LM and CM were performed to obtain pharmacokinetic parameters in healthy conditions. Binding parameters were obtained from an in vitro study using ultrafiltration. Simulation studies indicated that an increase of plasma AGP levels resulted in a decrease of both volume of distribution at steady state (Vdss) and total body clearance (Cltot) for all the drugs. Elimination rate constant for LM was almost unchanged by an increase of plasma AGP levels, whereas those for EM and CM were increased. Plasma concentration,time profiles at a high AGP level (often observed in pathophysiological conditions) were also simulated. All of the total plasma concentration,time profiles were different from those at normal AGP level. The differences were characterized by a higher initial concentration with faster or similar elimination. Unbound plasma concentration,time profile of LM was unaffected by AGP levels, whereas EM and CM were eliminated from plasma more rapidly at high AGP level. These results suggested that adjustment of dosage regimen of EM and CM is required in pathophysiological conditions, but that of LM is not required. [source] A New Method for Simultaneous Estimation of Micellization Parameters from Conductometric DataCHEMISTRY - A EUROPEAN JOURNAL, Issue 20 2004Nenad Jal, enjak Dr. Abstract A simple method for determination of the counterion binding parameter (,) and aggregation number (N) from conductivity data is proposed. The method is based on fitting the values of the first derivative of conductivity (,) versus total surfactant concentration (ct) function according to the equation derived from the mass action model (MAM) by using different conductivity models. Sodium dodecylsulphate (SDS) and dodecyltrimethylammonium bromide (DTAB) were chosen for validation of the proposed method. It was shown that the method gives a fairly accurate values for micellisation parameters of SDS (N=51,64, ,=0.74,0.75) and DTAB (N=56,62, ,=0.77,0.79), both in good agreement with the literature data. In addition, application of the proposed method does not require the value of the critical micelle concentration (cmc). [source] Study of the subunit interactions in myosin phosphatase by surface plasmon resonanceFEBS JOURNAL, Issue 6 2000Attila Tóth The interactions of the catalytic subunit of type 1 protein phosphatase (PP1c) and the N-terminal half (residues 1,511) of myosin phosphatase target subunit 1 (MYPT1) were studied. Biotinylated MYPT1 derivatives were immobilized on streptavidin-biosensor chips, and binding parameters with PP1c were determined by surface plasmon resonance (SPR). The affinity of binding of PP1c was: MYPT11,296 > MYPT11,38 > MYPT123,38. No binding was detected with MYPT11,34, suggesting a critical role for residues 35,38, i.e. the PP1c binding motif. Binding of residues 1,22 was inferred from: a higher affinity binding to PP1c for MYPT11,38 compared to MYPT123,38, as deduced from SPR kinetic data and ligand competition assays; and an activation of the myosin light chain phosphatase activity of PP1c by MYPT11,38, but not by MYPT123,38. Residues 40,296 (ankyrin repeats) in MYPT11,296 inhibited the phosphorylase phosphatase activity of PP1c (IC50 = 0.2 nm), whereas MYPT11,38, MYPT123,38 or MYPT11,34 were without effect. MYPT140,511, which alone did not bind to PP1c, showed facilitated binding to the complexes of PP1c,MYPT11,38 and PP1c,MYPT123,38. The inhibitory effect of MYPT140,511 on the phosphorylase phosphatase activity of PP1c also was increased in the presence of MYPT11,38. The binding of MYPT1304,511 to complexes of PP1c and MYPT11,38, or MYPT11,296, was detected by SPR. These results suggest that within the N-terminal half of MYPT1 there are at least four binding sites for PP1c. The essential interaction is with the PP1c-binding motif and the other interactions are facilitated in an ordered and cooperative manner. [source] Preparation and Characteristics of Esculin-Imprinted PolymersHELVETICA CHIMICA ACTA, Issue 6 2007Guo-Song Wang Abstract Four molecularly imprinted polymers (MIPs) were prepared in MeOH with esculin (=6,7-dihydroxycoumarin 6-(, - D -glucopyranoside)=6-(, - D -glucopyranosyloxy)-7-hydroxy-2H -1-benzopyran-2-one) as the imprinted molecule, methacrylic acid (=2-methylprop-2-enoic acid; MAA), acrylamide (=prop-2-enamide; AM), 4-vinylpyridine (=4-ethenylpyridine; 4-VP), or 2-vinylpyridine (=2-ethenylpyridine; 2-VP) as the functional monomer, respectively, as well as ethylene glycol dimethacrylate (=2-methylprop-2-enoic acid ethane-1,2-diyl ester; EGDMA) as the cross-linking agent. The interaction between the template and the functional monomers was investigated by fluorescence and UV spectrophotometry, respectively, which revealed the presence of esculin/monomer complexes in the stoichiometric ratio 1,:,2 in the pre-polymerization mixture. The resultant polymers were studied in equilibrium binding experiments to evaluate the recognition ability and the binding capacity towards esculin. The results showed that MIP1, prepared with MAA as the functional monomer, exhibited advantageous characteristics of high binding capacity, optimal imprinting effect, and good selectivity towards esculin. The Scatchard analysis indicated that there are two types of binding sites in MIP1, and its binding parameters including the apparent maximum numbers of binding sites and the dissociation constants were calculated. Finally, by packing an SPE column (SPE=solid-phase extraction) with MIP1, the esculin was separated and enriched successfully by this sorbent from samples of Cortex fraxini, and the average recovery was up to 74.7%. [source] A simple and convenient approach for evaluation of the parameters of ligand,receptor interaction.JOURNAL OF MOLECULAR RECOGNITION, Issue 2 2008Receptor blocking index, its application Abstract A new approach for determination of the parameters for ligand-receptor interaction, which is based on so-called dilution coordinates, was developed earlier. Equations that allow evaluation of not only the affinity of ligand-receptor interaction but also of the amount of free (or occupied by corresponding ligand) receptors were suggested. The most important advantage of this approach as compared with well-known methods is the ability to determine the binding parameters for ligand-receptor interaction even for the cases in which ligand and receptor are already present in a mixture and separation of counterparts from each other is technically difficult or even impossible. Due to this reason, the proposed approach can be especially useful for studying interactions between highly-labile biological receptors and corresponding ligands as found in vivo. In the present paper I continue to consider how to determine the binding parameters for a given ligand-receptor interaction if the value of receptor blocking index is determined experimentally. Copyright © 2008 John Wiley & Sons, Ltd. [source] Survey of the year 2004 commercial optical biosensor literatureJOURNAL OF MOLECULAR RECOGNITION, Issue 6 2005Rebecca L. Rich Abstract The year 2004 represents a milestone for the biosensor research community: in this year, over 1000 articles were published describing experiments performed using commercially available systems. The 1038 papers we found represent a ,10% increase over the past year and demonstrate that the implementation of biosensors continues to expand at a healthy pace. We evaluated the data presented in each paper and compiled a ,top 10' list. These 10 articles, which we recommend every biosensor user reads, describe well-performed kinetic, equilibrium and qualitative/screening studies, provide comparisons between binding parameters obtained from different biosensor users, as well as from biosensor- and solution-based interaction analyses, and summarize the cutting-edge applications of the technology. We also re-iterate some of the experimental pitfalls that lead to sub-optimal data and over-interpreted results. We are hopeful that the biosensor community, by applying the hints we outline, will obtain data on a par with that presented in the 10 spotlighted articles. This will ensure that the scientific community at large can be confident in the data we report from optical biosensors. Copyright © 2005 John Wiley & Sons, Ltd. [source] Determination of binding constants and stoichiometries for platinum anticancer drugs and serum transport proteins by capillary electrophoresis using the Hummel-Dreyer methodJOURNAL OF SEPARATION SCIENCE, JSS, Issue 2 2005Alexander V. Rudnev Abstract A CE method has been developed to evidence and quantitatively characterize the interaction between platinum-based antitumor drugs and human serum proteins. This method is a variant of affinity CE modified regarding both experimental setup and data treatment so as to measure the peaks (or vacancies) that correspond to the bound drug when it slowly binds to the protein. Using the formalism of the Hummel-Dreyer method and cisplatin and oxaliplatin as test compounds, a protocol for determining albumin and transferrin binding constants and stoichiometries, including (and distinguished by) 48 hours of incubation of the reaction mixture, was elaborated. Relative affinities of drugs toward different proteins in aqueous solution at physiological pH, chloride concentration, and temperature were compared in terms of overall binding constants and numbers of drug molecules attached to the protein. The results indicate that both platinum drugs bind to albumin more strongly than to transferrin, supporting the concept that the albumin fraction is a major drug supply route for chemotherapeutical needs. From a comparison with the binding parameters measured previously for cisplatin by other methods, conclusions were drawn about the validity of CE as a simple and convenient method for assaying protein-drug reactions with slow kinetics. [source] N -methyl-d-aspartate Receptor Responses Are Differentially Modulated by Noncompetitive Receptor Antagonists and Ethanol in Inbred Long-Sleep and Short-Sleep Mice: Behavior and ElectrophysiologyALCOHOLISM, Issue 12 2000Taleen Hanania Background: Short-sleep (SS) mice exhibit higher locomotor activity than do long-sleep (LS) mice when injected with low doses of ethanol or the noncompetitive N -methyl-D-aspartate receptor (NMDAR) antagonist MK-801 (dizocilpine). SS mice also have higher densities of brain NMDARs. However, two strains of LS X SS recombinant inbred (RI) mice also show differential activation to ethanol and MK-801, but have similar numbers of NMDARs. Here we used inbred LS (ILS) and SS (ISS) mice to investigate further the relationship between NMDARs and sensitivity to the stimulant effects of low doses of ethanol. Methods: Open field activity and spontaneous alternations were measured after saline or drug injection. [3H]MK-801 binding parameters were determined in hippocampus, cortex, striatum, and nucleus accumbens. Extracellular field excitatory postsynaptic potentials (fEPSPs) were recorded in the CA1 region of hippocampal slices. Results: Systemic injection of either ethanol or MK-801 increased locomotor activity to a greater extent in ISS mice than in ILS mice. The competitive NMDAR antagonist 2-carboxypiperazin-4-yl-propyl-1,1phosphonic acid (±CPP) depressed activity of ILS, but not ISS, mice. No strain differences were observed in spontaneous alternations or in the number or affinity of NMDARs in the brain regions examined. Likewise, the magnitudes of hippocampal NMDAR-mediated fEPSPs were similar in ILS and ISS mice and were inhibited to the same extent by a competitive NMDAR antagonist. However, both ethanol and the NMDAR NR2B receptor antagonist ifenprodil inhibited the late component of hippocampal NMDAR fEPSPs to a greater extent in ISS, than in ILS, mice. Conclusions: Differential ethanol- and MK-801-induced behavioral activation in ILS and ISS mice was not associated with differences in NMDAR number. Nonetheless, pharmacological differences in hippocampal NMDAR responsiveness suggest that ISS mice express NMDARs that have a greater sensitivity to noncompetitive, but not competitive, NMDAR antagonists. These differences, which may reflect differences in NMDAR subunit composition, could underlie the differential responsiveness to low doses of ethanol in ILS and ISS mice. [source] Dietary effects on insulin and glucagon plasma levels in rainbow trout (Oncorhynchus mykiss) and gilthead sea bream (Sparus aurata)AQUACULTURE NUTRITION, Issue 2 2009P. ROJAS Abstract The effects of dietary amino acid profile (based on muscle (M) or whole body composition (WB) and the balance between indispensable (IAA) and dispensable amino acids (DAA) in the diet, on plasma levels of insulin and glucagon, were analysed in rainbow trout and gilthead sea bream. Plasma insulin values (baseline and 6 h postfeeding) were higher in trout than in sea bream, but the relative postfeeding increase was more pronounced in sea bream. Within the same dietary amino acid profile, diets with lower IAA/DAA, had a lower effect on the postfeeding secretion of insulin in both species. Circulating levels of glucagon (baseline and postfeeding relative increases) were higher in sea bream. In trout, diets with WB amino acid profile had a greater secretory effect on postfeeding glucagon than did diets with M profile, while gilthead sea bream showed an inverse response to circulating glucagon with respect to diet. Muscle insulin and insulin growth factor-I binding parameters were not affected by the dietary regimen. The postfeeding glucagon response depends on both the dietary AA profile and the fish species, while that of insulin seems to be more uniform, and is affected in a similar way regardless of the species. [source] Nonstationary disposition of valproic acid during prolonged intravenous infusion: contributions of unbound clearance and protein bindingBIOPHARMACEUTICS AND DRUG DISPOSITION, Issue 6 2001Tori L. Arens Abstract Circadian variations in disposition have been observed for a variety of agents, including anticonvulsants. Valproic acid (VPA), an anticonvulsant used to control generalized and partial seizures, has exhibited diurnal oscillations in steady-state concentrations during long-term administration to humans and non-human primates. The present study was conducted to assess potential diurnal changes in the disposition of VPA during prolonged i.v. infusion in rats. Animals, maintained on a strict 12-h per day light cycle, were equipped with venous cannulae and an arterial microdialysis probe. VPA was administered as a 50-mg/kg loading dose followed by a 42 mg/kg/h infusion for 70 h. Blood and microdialysate samples were obtained at timed intervals after establishment of steady-state throughout two complete light/dark cycles; and total (serum) and unbound (microdialysate) VPA was determined by gas chromatography. Modest oscillations (6,7 h period) in total and unbound VPA were observed; clearance and binding parameters were not different between light and dark periods. However, unbound clearance increased, and unbound fraction decreased, with time over the course of the infusion. These results suggest that time-dependent changes in VPA disposition occur in rats, although oscillations in steady-state concentrations do not appear to be diurnal in nature. Copyright © 2001 John Wiley & Sons, Ltd. [source] Influence of N-Terminal Hydrophobicity of Cationic Peptides on Thermodynamics of their Interaction with Plasmid DNACHEMICAL BIOLOGY & DRUG DESIGN, Issue 5 2009Geetha N. Goparaju There is a need to understand the thermodynamics of interaction of cationic peptides with DNA to design better peptide based non-viral gene delivery vectors. The main aim of this study was to understand the influence of N-terminal hydrophobicity of cationic amphiphilic peptides on thermodynamics of interaction with plasmid DNA. The model peptides used were TATPTD and TATPTDs modified at the N-terminal with hydrophobic amino acids. The thermodynamic binding data from isothermal titration calorimetry were compared with ethidium bromide analysis and ultrafiltration to correlate the binding parameters with the structural features of the various peptides used. It was observed that peptides having a smaller hydrophobic domain at the N-terminal have good DNA condensing ability compared with the ones with a longer hydrophobic domain. Calorimetry of peptides that reached saturation binding indicated that enthalpy and entropy are favorable for the interaction. Moreover, the interaction of these peptides with DNA appears to be predominantly electrostatic. [source] | |||||||||||||