Modelling the Influence of Nanoparticles in the Phase Behaviour of an Epoxy/Polystyrene Mixture, 2

The influences of nanoparticle size and concentration on the thermodynamic behaviour of epoxy/polystyrene blends are evaluated in the framework of Ginzburg's simple analytical theory. Two approaches have been employed: NPEPO (for particles coated with epoxy groups) and NPFEN (for particles coated with phenyl groups). Using NPEPO, the particles are found to prefer the phase richer in epoxy, whereas the opposite occurs for NPFEN. The particles size significantly influences blend compatibility. When the particle radius Rp is about the same size as the radius of gyration R g of PS, the compatibility of blends increases with particle concentration, whereas for R p > R g , higher particle concent…

Incorporation of Silica Nanospherical Particles into Epoxy-Amine Crosslinked Materials

Composites of silica nanospheres coated with crosslinked epoxy–amine were synthesised and examined by 29Si-magic-angle-spinning nuclear magnetic resonance spectroscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy and scanning electron microscopy. The most representative fact is that epoxy-modified nanospheres lost less weight at high temperatures. At temperatures greater than 300°C the loss of weight for epoxy-modified nanospheres was rather lower than for unmodified nanospheres. This helped them to retain their structures, as the loss of weight can have adverse effects on network defects, due to the loss of crosslinks by unit of volume.

Miscibility of a DGEBA Based Epoxy Resin Blended with Thermoplastic Mixtures of Poly(styrene) and Block Copolymers: Influence of the Copolymer Content and Chemical Nature

The miscibility and phase behavior of ternary systems formed by a DGEBA based epoxy resin, polystyrene (PS), and diblock copolymers has been investigated through phase diagrams experimentally obtai...

Separation Efficiency of Two SEC Packings: Comparison of Chromatographic, Thermodynamic, and Fractal Parameters

Abstract The size‐exclusion chromatographic (SEC), thermodynamic, and fractal behavior of two organic packings, named TSK‐Gel HHR and TSK‐Gel HXL, has been compared in order to assess their separation efficiency. The different experimental elution trends of five solvent/polymer systems have been evidenced through the universal calibration curves, and the existence of secondary mechanisms (such as adsorption) has been quantified by the chromatographic partition coefficient K p. From a thermodynamic point of view, the swelling and crosslinking degrees have been evaluated and compared in the two gels assayed. Moreover, the values of some fractal parameters (fractal dimension of the surface and…

Size‐Exclusion Chromatography in Single and Mixed Eluents: Effects of Sorption of Solvent and Polymer Solutes on Novel TSK‐Gel Super AW Columns

Abstract The novel TSK Gel Super AW column packing is based on a polymeric gel with residual hydrophobic and hydrophilic groups on the gel surface. These groups can act as active sites making the chromatographic support compatible both with polar and apolar eluents. This interesting property allows the elution of systems of very different polarities but also induces binary polymer–gel, polymer–solvent, and solvent–gel enthalpic interactions that lead to a mixture of separation mechanisms: repulsion, partition, or adsorption besides the main size‐exclusion chromatography (SEC) mechanism. For this reason, the elution behaviour of many and diverse polymer/solvent systems has been studied in th…

Stabilisation of mixed peptide/lipid complexes in selective antifungal hexapeptides

AbstractThe design of antimicrobial peptides could have benefited from structural studies of known peptides having specific activity against targetmicrobes, but not toward other microorganisms. We have previously reported the identification of a series of peptides (PAF-series) activeagainst certain postharvest fungal phytopathogens, and devoid of toxicity towards E. coli and S. cerevisiae [Lo´pez-Garci´a et al. Appl.Environ. Microbiol. 68 (2002) 2453]. The peptides inhibited the conidia germination and hyphal growth. Here, we present a comparativestructural characterisation of selected PAF peptides obtained by single-amino-acid replacement, which differ in biological activity. Thepeptides w…

The amphipathic peptide mellitin as a tool to study the membrane-dependent activation of tissue transglutaminase

The role of membrane phospholipids on the cross-linking activity of guinea pig liver (tissue) transglutaminase has been investigated using the amphipathic model peptide melittin as glutaminyl substrate and the primary amine monodansylcadaverine as extrinsic amine donor. A marked increase of transglutaminase catalytic activity was observedin vitro assays in the presence of neutral membrane phospholipids. In contrast, activation was abolished in the presence of membranes containing pure anionic lipids. Enzyme activation could be ascribed to a direct binding of the lipid to the protein as demonstrated in enzymatic assays using a non membrane-interacting peptide (Cbz-Gln-Gly). The data obtained…

Macromolecules in ordered media. 6. Effect of the phospholipidic bilayer composition on poly (2-vinyl pyridine) - liposome association

The intrinsic fluorescence emission of poly (2-vinyl pyridine) has been exploited in order to gain insight into the nature of the interaction with artificial membranes. In this contribution, the study has been focused on the effect of the phospholipid bilayer composition on the association to the polymer. Fluorescence data have been converted into association isotherms and then analyzed in terms of both binding and partition models. Results show that the association is enhanced by the presence of negative charged phospholipid as a constituent of the liposomal membrane. The strong surface density dependence of the polymer-liposome association indicatesthe predominantly electrostatic nature o…

A thermodynamic approach to study hydrogen-bonding interactions in solvent/solvent/polymer ternary systems

Abstract A thermodynamic approach based on both the classical Flory–Huggins (FH) formalism and the association equilibria (AE) theory has been developed to study the solubility properties of a system formed by a proton-donor solvent (A), a proton-acceptor solvent (B) and a proton-acceptor polymer (C). The miscibility of this ternary system is attained by competitive specific interactions via hydrogen-bonding established between the hydroxyl and carbonyl interacting groups of either solvent–solvent (AB) or solvent–polymer (AC) system components. The binary AB and AC specific interactions and their dependence with the system composition as well as with the extent of the association equilibriu…

Characterization of gramicidin A in an inverted micellar environment. A combined high-performance liquid chromatographic and spectroscopic study

We have investigated the conformational adaptability of gramicidin A incorporated into reverse micelles of sodium bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane/water, a so far unexplored "host" membrane-mimetic model system for this peptide. A high-performance liquid chromatographic strategy previously developed for the study of gramicidin in phospholipid vesicles and normal micelles [Bañó et al. (1989) FEBS Lett. 250, 67; Bañó et al. (1991) Biochemistry 30, 886] has been successfully extended to this system. The method has permitted the separation of peptide conformational species, namely, double-stranded dimers and monomers, and an accurate quantitation of their proportion in the invert…

Tyrosinase Enzyme: 1. An Overview on a Pharmacological Target

The tyrosinase enzyme (EC 1.14.18.1) is an oxidoreductase inside the general enzyme classification and is involved in the oxidation and reduction process in the epidermis. These chemical reactions that the enzyme catalyzes are of principal importance in the melanogenesis process. This process of melanogenesis is related to the melanin formation, a heteropolymer of indolic nature that provides the different tonalities in the skin and helps to the protection from the ultraviolet radiation. However, a pigment overproduction, come up by the action of the tyrosinase, can cause different disorders in the skin related to the hyperpigmentation. Several studies mainly focused on the characteristics …

Modelling Monomer/Disc Composites Phase Behaviour

Summary: A model developed by Balazs' group to explain the phase behaviour of polymer/clay composites is extended to obtain an expression for the free energy of polymer/thin disc mixtures. Phase diagrams for monomer/disc mixtures are built by minimizing the free energy and calculating the chemical potentials of the three system components. Via the comparison of the diagrams, it is studied the effects of nanodisc size and interaction parameters on mixture stability and attained morphology. The performed predictions between monomers and discs give criteria that advance the properties of the mixture. Changes in monomer concentration and interaction parameters provide a means to prevent van der…

Membrane Lipid-Polypeptide Molecular Associations in Non Aqueous Solvent. Effect of Phosphatidylcholine Concentration and Temperature and their Influence on the Gramicidin a Dimer-Monomer Conformational Equilibrium

Abstract The use of an Ultrastyragel 500 A column for the study of interactions between phosphatidylcholine and gramicidin A in tetra-hydrofuran is described. Analysis of vacant peak has allowed to establish the influence that eluent lipid composition, concentration of injected gramicidin and temperature have on the interaction. At 20[ddot] C, for the assayed phospholipid concentration range (0.04 to 0.14 %, w/v), the lipid/polypeptide molar binding ratio, BR, varies from 1.3 to 6.9. An increase in temperature from 20 to 40[ddot] C causes a decrease in BR of about 20 %. On the other hand, the interaction releases some of the water bound to the lipid polar head, suggesting that the binding i…

Incorporation of Silica Nanospherical Particles in Epoxy–Amine Crosslinked Materials II. Dynamic Mechanical Measurements of Epoxy Matrix-Silica Nanocomposites

The mechanical strengths of epoxy composites reinforced with silica nanospheres, unfunctionalised or functionalised with either amine or epoxy groups, increase up to a proportion of 5 wt.% of filler, as reflected in a study of the shear storage modulus carried out in dynamic mechanical analysis. This improvement is observed in both glassy and rubbery states, moderately affecting the glass transition temperature of the material. From this percentage of strengthening substance the mechanical properties begin to deteriorate, but keeping (up to 10 wt.% of strengthening material), a greater storage modulus in shear than that of the pristine epoxy resin. A trend can be discerned as the percentag…

Modelling the hydrogen-bonding interactions in a copolymer/biodegradable homopolymer blend through excess functions

A recent theoretical approach based on the coupling of both the Flory-Huggins (FH) and the Association Equilibria thermodynamic (AET) theories was modified and adapted to study the miscibility properties of a multi- component system formed by two polymers (a proton-donor and a proton-acceptor) and a proton-acceptor solvent, named copolymer(A)/solvent(B)/polymer(C). Compatibility between polymers was mainly attained by hydrogen- bonding between the hydroxyl group on the phenol unit of the poly(styrene-co-vinyl phenol) (PSVPh) and the car- bonyl group of the biodegradable and environmentally friendly poly(3-hydroxybutyrate) (PHB). However, the self- association of PSVPh and specific interacti…

Ternary Polymer Solutions with Hydrogen Bonds, 1

The Flory Huggins methodology coupled to AET has been extended to ternary polymer systems, in particular to solvent (A)/polymer 1 (B)/polymer 2 (C) systems, with the two polymers displaying H-bonding interactions. Because the H-bonding can perturb the randomness of polymeric conformations, the change in Gibbs free energy of mixing, AG, should arise from changes in combinatorial entropy as well as in interaction energy. The combinatorial part of AG is evaluated through AET as a function of the association constant η between B and C components, the autoassociation constant a between B components, and the independent number m of interaction sites of acceptor C. The enthalpic contribution is ev…

The Fractal Approach to Secondary Mechanisms in SEC

Abstract Size‐exclusion chromatography (SEC) is one of the most used experimental techniques to characterize polymers in solution; it has been applied to interpret the elution behaviour of many polymer‐solvent systems in five types of column packings. The experimental data have revealed that the classical universal calibration is not accomplished. Deviations from a unique curve are a consequence of two effects: entropic (exclusion by size) and enthalpic (binary and ternary interactions between solvent, polymer, and gel), which results in secondary mechanisms accompanying the main “ideal” SEC separation mechanism. Therefore, three approaches of building a calibration curve have been compared…

Size-Exclusion Chromatographic Determination of Polymer Molar Mass Averages Using a Fractal Calibration

The characterization of polymers by size-exclusion chromatography basically consists of the determination of the weight-average molar mass (Mw), number-average molar mass (Mn), and polydispersity index (I). An accurate estimation of these magnitudes requires the use of a reliable and trusted calibration curve. Three procedures for building up a calibration curve are analyzed in this work. The first is the classical universal calibration (UC), based on the elution of tetrahydrofuran-polystyrene in a system as reference. The second is based on the proper calibration curve made with standards of the sample under study. However, two main drawbacks arise when using these methodologies: the nonfu…

Time-dependent monomerization of gramicidin A, enhanced by phosphatidylcholine in non-polar solvents

Abstract The usefulness of size-exclusion high-performance liquid chromatography for the study of gramicidin A dimer—monomer conformational equilibrium in non

The Closed/Open Model for Lipase Activation. Addressing Intermediate Active Forms of Fungal Enzymes by Trapping of Conformers in Water-Restricted Environments

The behavior of prototypic fungal lipases in a water-restricted environment has been investigated by exploiting the reported experimental strategy that allows the trapping (freeze-drying) of the enzyme in the conformation present in aqueous solution and to subsequently assay it in nonaqueous media [Mingarro, I., Abad, C., and Braco, L. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 3308-3312]. We now report, using simple esterification as well as acidolysis (triglycerides as substrates) as nonaqueous model reactions, that the presence of a detergent (n-octyl-beta-glucopyranoside) in the freeze-drying buffer, at concentrations below the critical micellar concentration, generates different catalyti…

The fractal calibration method applied to the characterization of polymers in solvent mixtures and in mixed gel packings by SEC.

The size-exclusion chromatographic (SEC) behaviour of different solvent/polymer systems in three packing sets has been analysed from fractal considerations. The three-column sets studied are specifically formed by: (i) 'pure' micro-styragel, (ii) 'mixed' TSK Gel H(HR + XL + HR) and (iii) mixed TSK Gel H(XL + HR + XL). The experimental data reveals that in most of the systems assayed the classical universal calibration (UC) is not fulfilled, denoting the existence of secondary effects accompanying the main SEC mechanism. In order to obtain an accurate characterization of different polymers eluted in solvent mixtures and/or mixed packings, the use of a reliable and trusted calibration curve i…

Solution properties of polyelectrolytes

Abstract Chromatographic data for sodium polystyrene sulphonate were obtained on both silica- and polymer-based size-exclusion supports using mobile phases of various pH and ionic strength. Deviations of the elution volume were observed towards both lower and higher values relative to the reference calibration graph obtained with uncharged standards. An empirical correlation is proposed in order to account for all the secondary effects observed. The general applicability of this correlation was further tested for chromatographic data obtained for a series of peptides and proteins on a silica-based support under very different eluent conditions. Deviations from ideal elution behaviour such a…

Preferential solvation of poly(methyl methacrylate) and a bisphenol A diglycidyl ether by size-exclusion chromatography

The preferential adsorption coefficient, lambda, of poly(methyl methacrylate), PMMA, in solutions formed by an epoxy resin in tetrahydrofuran (THF), was studied by size-exclusion chromatography (SEC). Only PMMA of lowest molar mass was preferentially solvated by epoxy but at low concentrations of epoxy in the mixture. At higher epoxy content PMMA was preferentially solvated by THF. A simultaneous and competitive solvation between the specific interactions PMMA-epoxy and the self association of epoxy at high concentrations would be the responsible of this inversion point. The more compacted coil of PMMA of higher molecular weights in solution could explain the lack of interaction of these po…

Addressing substrate glutamine requirements for tissue transglutaminase using substance P analogues

AbstractWe have investigated the effect on the substrate requirements for guinea pig liver (tissue) transglutaminase of a set of 11 synthetic glutamine substitution analogues making up the full sequence of the naturally occurring tissue transglutaminase substrate substance P. While a number of peptide sequences derived from proteins that are well-recognized as tissue transglutaminase substrates have been studied, the enzyme activity using substitution analogues of full-length natural substrates has not been investigated as thoroughly. Thus, our set of substance P analogues only differs from one to other by one amino acid mutation while the length (of the peptide) is maintained as in the nat…

Ligand-based discovery of novel trypanosomicidal drug-like compounds: In silico identification and experimental support

Abstract Two-dimensional bond-based linear indices and linear discriminant analysis are used in this report to perform a quantitative structure–activity relationship study to identify new trypanosomicidal compounds. A database with 143 anti-trypanosomal and 297 compounds having other clinical uses, are utilized to develop the theoretical models. The best discriminant models computed using bond-based linear indices provides accuracies greater than 90 for both training and test sets. Our models identify as anti-trypanosomals five out of nine compounds of a set of already-synthesized substances. The in vitro anti-trypanosomal activity of this set against epimastigote forms of Trypanosoma cruzi…

Thermodynamic study on phase equilibrium of epoxy resin/thermoplastic blends

Abstract The experimental phase diagrams (cloud point curves) of three series of epoxy/thermoplastic blends, namely, epoxy/polystyrene (PS), epoxy/poly(ether sulfone) (PES), and epoxy/poly(ether imide) (PEI) as a function of molar mass and composition have been analysed from a thermodynamic point of view. A model based on the Flory–Huggins lattice theory considering the concentration dependence of the interaction parameter as predicted by Koningsveld was employed to determine the equilibrium compositions, and concentration and temperature dependent interaction parameters. Binodal, spinodal, and critical point data have been computed and show good agreement with experimental data.

Selective labelling of melittin with a fluorescent dansylcadaverine probe using guinea-pig liver transglutaminase

Abstract Melittin, a C-terminal peptide, incorporated the fluorescent probe monodansylcadaverine (DNC) when catalysed by guinea-pig liver transglutaminase and Ca2+, as determined by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). A 1:1 adduct DNC-melittin was identified in which a single glutamine residue out of two, i.e. Gln25, acts as acyl donor. Incubation of melittin with transglutaminase in the absence of DNC originated high molecular mass complexes indicative that the peptide lysine residue can act as an acyl acceptor. The DNC-melittin was about 3 times more active in the lysis of red cell membranes than native melittin. Fluorescence study of the lab…

Interaction of quinine with negatively charged lipid vesicles studied by fluorescence spectroscopy Influence of the pH

Abstract The interaction of quinine with dimyristoylphosphatidic acid (DMPA) and dimyristoylphosphatidyl glycerol (DMPG) small unilamellar vesicles in the gel phase was studied by steady-state fluorescence spectroscopy at pHs 7, 6, 5 and 4 and 20°C. In aqueous solution, with excitation at 335 nm, the emission fluorescence spectrum of quinine varied with pH reflecting the occurrence of different charged species of the drug. In all cases, the emission maximum centered at 383 or 443 nm shifted to lower wavelength in the presence of vesicles. This indicates that the membrane-bound state quinine is in an environment of low polarity. Drug monocationic species were deeply buried in DMPG relative t…

Cover Picture: Macromol. Theory Simul. 1/2007

New high-performance liquid chromatography-based methodology for monitoring the conformational transitions of self-associating hydrophobic peptides, incorporated into liposomes.

A new high-performance size-exclusion chromatographic strategy is reported for the analysis of the hydrophobic self-associating peptide gramicidin A, incorporated into artificial phospholipid vesicles (liposomes). The method is based on the direct injection of a few microlitres of the gramicidin A-containing liposome suspension into the column, which is eluted with a non-polar solvent, such as tetrahydrofuran. The type and amount of information which can be derived from this methodology have been evaluated. Using this chromatographic approach, a correlation has been unambiguously shown to exist between the organization of the peptide in the vesicles and a number of variables involved in the…

Prediction of Aquatic Toxicity of Benzene Derivatives to Tetrahymena pyriformis According to OECD Principles

Background: Many QSAR studies have been developed to predict acute toxicity over several biomarkers like Pimephales promelas, Daphnia magna and Tetrahymena pyriformis. Regardless of the progress made in this field there are still some gaps to be resolved such as the prediction of aquatic toxicity over the protozoan T. pyriformis still lack a QSAR study focused in accomplish the OECD principles. Methods: Atom-based quadratic indices are used to obtain quantitative structure-activity relationship (QSAR) models for the prediction of aquatic toxicity. Our models agree with the principles required by the OECD for QSAR models to regulatory purposes. The database employed consists of 392 substitut…

Influence of the Copolymer Content on the Miscibility, Phase Behaviour and Morphology of a DGEBA/Polystyrene/Styrene-co-Maleic Anhydride Ternary Blend

The miscibility, phase behaviour and morphology of ternary systems, formed by a DGEBA-based epoxy resin, polystyrene (PS) and styrene-co-maleic anhydride (SMA) copolymers have been investigated through phase diagrams. The analysis has been done in the absence and in the presence of 4,4'-methylene bis(2,6-diethyl aniline) (MDEA) as curing agent. In both cases, the influence of the copolymer content on blend compatibility, has been discussed. The results show that miscibility is enhanced by the presence of SMA copolymer in the blend and as the MA content in the copolymer is increased (from 7 to 14wt.-%), due to specific H-bonding interactions between the H-donor hydroxyl groups on the epoxy r…

Dynamic Mechanical Measurements of Epoxy Matrix-Silica Nanocomposites II

The mechanical properties of epoxy-silica nanocomposites have been studied; the silica nanosphere fillers used were un-functionalised, functionalised with amine, with epoxy, or a mixture of both kinds. Dynamic mechanical analysis measurements revealed an increase in the shear storage modulus, for all samples with a filler content of 3–5%. Improvements were observed in the glassy and rubbery states, without affecting the glass transition temperature of the materials. Above these strengthening percentages, the mechanical properties began to deteriorate, but in all cases they remained superior to those of the pristine epoxy resin. For low strengthening percentages, samples reinforced with bot…

Structural characterisation of the natural membrane-bound state of melittin: a fluorescence study of a dansylated analogue

Abstract The binding of a dansylated analogue of melittin (DNC–melittin) to natural membranes is described. The cytolytic peptide from honey bee venom melittin was enzymatically labelled in its glutamine-25 with the fluorescent probe monodansylcadaverine using guinea pig liver transglutaminase. The labelled peptide was characterised functionally in cytolytic assays, and spectroscopically by circular dichroism and fluorescence. The behaviour of DNC–melittin was, in all respects, indistinguishable from that of the naturally occurring peptide. We used resonance energy transfer to measure the state of aggregation of melittin on the membrane plane in synthetic and natural lipid bilayers. When bo…

HPLC in the characterisation of conformational species of linear gramicidins

Abstract High-performance size-exclusion chromatography (HPSEC) has proved to be a highly simplified and rapid procedure to investigate the conformational behaviour of gramicidin A inserted in different model membrane systems (lipid dispersions, liposomes, micelles) based on the separation of double-stranded dimers and monomers present in the lipid assemblies. The HPSEC approach has been extended to the characterisation of acylated analogs of gramicidin (palmitoyl- and oleoyl-derivatives) and a series of peptide analogs where tryptophan residues were chemically modified or replaced by less polar residues (phenylalanine or naphtylalanine) lacking H-bonding ability. The Chromatographic result…

An Analysis of the Concentration Effects on Elution Volumes by Using the Preferential Solvation Parameter in Two SEC Packings

Size exclusion chromatography (SEC) elution behavior of different solvent/polymer systems has been analyzed from chromatographic, thermodynamic, and hydrodynamic points of view in two organic column packings based on polystyrene/divinylbenzene (PS/DVB) copolymer, μ-styragel and TSK-Gel H HR . Although both packings present similar chromatographic properties, some differences arise when eluting the same systems. The values of the adsorption distribution coefficients, K p , have been correlated with the preferential solvation coefficient, λ (see Figure), for both packings, showing that those systems with lower and negative λ values (denoting higher preferential solvation of the polymer by the…

Environment- and sequence-dependent modulation of the double-stranded to single-stranded conformational transition of gramicidin A in membranes.

The role of the membrane lipid composition and the individual Trp residues in the conformational rearrangement of gramicidin A along the folding pathway to its channel conformation has been examined in phospholipid bilayers by means of previously described size-exclusion high-performance liquid chromatography HPLC-based strategy (Bano et al. (1991) Biochemistry 30, 886). It has been demonstrated that the chemical composition of the membrane influences the transition rate of the peptide rearrangement from double-stranded dimers to beta-helical monomers. The chemical modification of Trp residues, or its substitution by the more hydrophobic residues phenylalanine or naphthylalanine, stabilized…

Comparative evaluation of the swelling and degrees of cross-linking in three organic gel packings for SEC through some geometric parameters.

Abstract The size exclusion chromatographic (SEC) behavior of five solvent/polymer systems in three organic column packings based on polystyrene/divinylbenzene (PS/DVB) copolymer, TSK-Gel H HR , μ-styragel and TSK-Gel H XL , has been compared. All the packings offer similar characteristics (pore size, particle size and efficiency) but some differences have been found when eluting the same systems. The different elution behavior observed in both polymeric gels has been analyzed in terms of their swelling and cross-linking degrees and of the fractal parameters. From the Universal Calibration plots, values of the chromatographic partition coefficient, K p , have been obtained and using some eq…

Interaction of Quinine with Model Lipid Membranes of Different Compositions

The binding of a drug such as Quinine with neutral and negatively charged small unilamellar lipid vesicles at pH 7 and 37 degrees C was investigated. Changes in the fluorescence properties of the drug after association with the liposomes were used to obtain binding isotherms over a range of phospholipid compositions at different ionic strengths. Under such conditions, the association was strongly enhanced by the negative net charge of the bilayer but diminished by the increasing presence of electrolytes in the aqueous media. Binding data were analyzed in terms of a surface partition equilibrium without and after correcting for electrostatic effects by means of the Gouy-Chapman theory. The i…

Compatibility between polystyrene copolymers and polymers in solution via hydrogen bonding

Abstract It has been applied the concept of improving miscibility, by introducing and optimizing the extent of intermolecular hydrogen-bonding interactions between two polymers. We select a commodity polymer such as polystyrene, to study the compatibility in chloroform with poly(vinyl pyridine) and poly(vinyl pyrrolidone), both considered as proton acceptors. In order to enhance polymer–polymer miscibility, polystyrene is slightly modified by copolymerization with methacrylic acid, in the first case, and with vinyl-phenol comonomer, in the second one. In this way, two series of polystyrene-based copolymers are synthesized and characterized bearing ca. 8% (w/w) of –OH groups. The miscibility…

Size-exclusion high-performance liquid chromatography in the study of the autoassociating antibiotic gramicidin A in micellar milieu.

Gramicidin A (gA) is a polypeptide antibiotic which forms dimeric channels specific for monovalent cations in biological membranes. It is a polymorphic molecule that adopts several different conformations, double-stranded (ds) helical dimers (pore conformation) and single-stranded beta-helical dimers (channel conformation). This study investigated the conformational adaptability of gramicidin A when incorporated into micelles as membrane-mimetic model system. Taking advantage of our reported, versatile, size-exclusion high-performance liquid chromatography (SE-HPLC) strategy that allows the separation of double-stranded dimers and monomers, we have quantitatively characterized the conformat…

QSAR models for tyrosinase inhibitory activity description applying modern statistical classification techniques: A comparative study

Abstract Cluster analysis (CA), Linear and Quadratic Discriminant Analysis (L(Q)DA), Binary Logistic Regression (BLR) and Classification Tree (CT) are applied on two datasets for description of tyrosinase inhibitory activity from molecular structures. The first set included 701 tyrosinase inhibitors (TI) that are used for performance of inhibitory and non-inhibitory activity and the second one is for potency estimation of active compounds. 2D TOMOCOMD-CARDD atom-based quadratic indices are computed as molecular descriptors. CA is used to “rational” design of training (TS) and prediction set (PS) but it shows of not being adequate as classification technique. On the first data, the overall a…

Association Equilibria Theory for Polymers in Mixed Solvents with Specific Interactions

Hydrogen bonding plays an important role in determining the physical and thermodynamic properties of polar fluids. Theoretical and experimental aspects of polymers, in mixed solvents with hydrogen-bonding-specific interactions, are investigated using a simple association model based on the theory of association equilibria developed by Pouchly et al. The thermodynamic non-idealities are accounted for using a modified Flory-Huggins theory with effective g ij parameters. The entropic term of the above formalism has been formulated taking into account the pplymer segment-solvent as well as the solvent-solvent hydrogen-bond formation. Four equilibrium constants are introduced to make a realistic…

Binding of basic amphipathic peptides to neutral phospholipid membranes: a thermodynamic study applied to dansyl-labeled melittin and substance P analogues.

A thermodynamic approach is proposed to quantitatively analyze the binding isotherms of peptides to model membranes as a function of one adjustable parameter, the actual peptide charge in solution z(p)+. The main features of this approach are a theoretical expression for the partition coefficient calculated from the molar free energies of the peptide in the aqueous and lipid phases, an equation proposed by S. Stankowski [(1991) Biophysical Journal, Vol. 60, p. 341] to evaluate the activity coefficient of the peptide in the lipid phase, and the Debye-Huckel equation that quantifies the activity coefficient of the peptide in the aqueous phase. To assess the validity of this approach we have s…

Bond-based bilinear indices for computational discovery of novel trypanosomicidal drug-like compounds through virtual screening

Two-dimensional bond-based bilinear indices and linear discriminant analysis are used in this report to perform a quantitative structure-activity relationship study to identify new trypanosomicidal compounds. A data set of 440 organic chemicals, 143 with antitrypanosomal activity and 297 having other clinical uses, is used to develop the theoretical models. Two discriminant models, computed using bond-based bilinear indices, are developed and both show accuracies higher than 86% for training and test sets. The stochastic model correctly indentifies nine out of eleven compounds of a set of organic chemicals obtained from our synthetic collaborators. The in vitro antitrypanosomal activity of …

Multi-output Model with Box-Jenkins Operators of Quadratic Indices for Prediction of Malaria and Cancer Inhibitors Targeting Ubiquitin- Proteasome Pathway (UPP) Proteins.

The ubiquitin-proteasome pathway (UPP) is the primary degradation system of short-lived regulatory proteins. Cellular processes such as the cell cycle, signal transduction, gene expression, DNA repair and apoptosis are regulated by this UPP and dysfunctions in this system have important implications in the development of cancer, neurodegenerative, cardiac and other human pathologies. UPP seems also to be very important in the function of eukaryote cells of the human parasites like Plasmodium falciparum, the causal agent of the neglected disease Malaria. Hence, the UPP could be considered as an attractive target for the development of compounds with Anti-Malarial or Anti-cancer properties. R…

Fractal calibration in size-exclusion chromatography

Abstract The elution behaviour of different polymer–solvent systems in three types of organic columns for SEC has been compared and interpreted. The experimental data show that the classical universal calibration is not accomplished. Deviations from a unique curve are observed due to the binary and ternary interactions between the components of the system (solvent, polymer and gel) which results on secondary mechanisms accompanying the main pure or “ideal” SEC separation mechanism. Both, enthalpic and entropic effects are interpreted in terms of the swelling and crosslinking degrees of the gel packings, and are also related with the fractal characteristics of their surfaces, such as the fra…

Binding of a fluorescent dansylcadaverine-substance P analogue to negatively charged phospholipid membranes.

Abstract We have investigated the binding of a new dansylcadaverine derivative of substance P (DNC-SP) with negatively charged small unilamellar vesicles composed of a mixture of phosphatidylcholine (PC) and either phosphatidylglycerol (PG) or phosphatidylserine (PS) using fluorescence spectroscopic techniques. The changes in fluorescence properties were used to obtain association isotherms at variable membrane negative charges and at different ionic strengths. The experimental association isotherms were analyzed using two binding approaches: (i) the Langmuir adsorption isotherm and the partition equilibrium model, that neglect the activity coefficients; and (ii) the partition equilibrium m…

Study and comparison of interaction parameters and phase behavior of epoxy/polystyrene and epoxies copolymer polystyrene-b-poly(methyl methacrylate) blends

Thermodynamic studies in terms of phase separation behavior and interaction parameters, in mixtures of epoxy resins (thermoset materials) with thermoplastic additives, consisting of polystyrene (PS) or polystyrene-b-poly(methyl methacrylate) P(S-MMA) (with small proportion of MMA), have been carried out. It can be confirmed that the solubility of P(S-MMA) is an improvement on that of PS, with the prepolymer epoxy resins. Moreover, the interaction parameters are positive at temperatures in which phase separation is observed, but negative at temperatures in which the phase separation window disappears, in the corresponding phase diagram. POLYM. COMPOS., 2008. © 2008 Society of Plastics Engine…

Modelling the influence of nanoparticles in the phase behaviour of an epoxy/polystyrene mixture

Abstract The cloud point temperatures of four series of epoxy/polystyrene blends have been experimentally determined as a function of polymer mass and system composition. The phase diagrams show an UCST behaviour, increasing incompatibility as the molar mass increases. The Flory–Huggins theory with a concentration-dependent interaction parameter has been developed to study the compatibility of two polymers in presence of spherical nanoparticles. This theory has been first compared with the experimental cloud point curve in absence of nanoparticles, and secondly it has been used to predict the thermodynamic behaviour in presence of different volume fraction of nanoparticles. Nanoparticles co…

Treatment of poly(styrene-co-methacrylic acid)/poly(4-vinylpyridine) blends in solution under liquid–liquid phase-separation conditions. A new method for phase-separation data attainment from viscosity measurements

Phase diagrams are contributed for polymer mixture systems in solution. One polymer has proton-acceptor character and the other has growing proton-donor nature, which is reflected in the phase diagrams. Usually, these diagrams are obtained from size-exclusion chromatographic (SEC) measurements. A totally novel application, which is exposed in this report, is the construction of the phase diagram from the viscometric experiments of polymer mixtures. The evaluated binodal or cloud-point isotherms so built agree well with those from SEC. The results indicate an augmentation in the dimensions of donor polymer B, in the presence of acceptor polymer C, intensifying with the concentration of C, wh…

HPLC study on the ‘history’ dependence of gramicidin A conformation in phospholipid model membranes

AbstractA novel HPLC methodology for the study of gramicidin A reconstituted in model membranes has been tested in comparison with circular dichroism data. It is shown that this chromatographic technique not only corroborates most of the recent spectroscopic results but allows one to explain them in terms of mass fractions of different actual conformational species of GA in the phospholipid assemblies. In particular, the dependence of the inserted peptide configuration on the organic solvent and other parameters involved in the ‘history’ of the sample preparation and handling has been analyzed by HPLC in two phospholipid model systems: small unilamellar vesicles and micelles. Moreover, a sl…

Solution properties of polyelectrolytes. III. Effect of sodium polystyrene sulfonate concentration on viscometric and size exclusion chromatographic behavior at different ionic strengths

The effect of sodium polystyrene sulfonate concentration on the shape of its molecules in aqueous solution at different NaNO3 ionic strengths has been analyzed by viscometry and size-exclusion chromatography (s.e.c). An equation has been developed which predicts the intrinsic viscosity, [η] p.cp.cs, at finite concentration of both polyion (cp) and electrolyte (cs). The experimental results obtained by both techniques can be accounted for in terms of the theory. Several factors involved in the elution mechanism have been considered and the variation of the slopes of s.e.c. calibration curves with cp and cs has been discussed in terms of polyion conformation changes.

Negatively cooperative binding of melittin to neutral phospholipid vesicles

Abstract The association of basic amphipathic peptides to neutral phospholipid membranes is investigated in terms of binding and partition models. The binding of native and modified melittin to egg-yolk phosphatidylcholine vesicles is studied by steady-state fluorescence spectroscopy. The effect of the ionic strength shows an enhancement of the association as the ionic strength increases. After correction for electrostatic effects by the Gouy–Chapman theory, the melittin binding isotherms could be described by a partition model. In terms of conventional binding mechanisms, which do not take into account electrostatic effects, this would correspond to a negative cooperativity. A plausible wa…

Ternary Polymer Solutions with Hydrogen Bonds, 2

Experimental ternary phase diagrams for ternary systems CHL/PS-MAA/PVPy with diverse MAA contents have been determined by GPC. The presence of MAA in the copolymers gives rise to specific interactions, by hydrogen bond formation between both polymeric components, so strong that the isotherm for the system with the highest MAA content so far studied, CHL/PS-MAA(8%)/PVPy, is representative of a complex coacervation situation. By applying the theoretical background deduced by coupling the Flory-Huggins lattice model to the AET developed for ternary polymeric systems SPP with specific intermolecular interactions (via hydrogen bonds), free energy surfaces for the CHL/PS-MAA/PVPy systems are cons…

Interfacial activation-based molecular bioimprinting of lipolytic enzymes

Interfacial activation-based molecular (bio)-imprinting (IAMI) has been developed to rationally improve the performance of lipolytic enzymes in nonaqueous environments. The strategy combinedly exploits (i) the known dramatic enhancement of the protein conformational rigidity in a water-restricted milieu and (ii) the reported conformational changes associated with the activation of these enzymes at lipid-water interfaces, which basically involves an increased substrate accessibility to the active site and/or an induction of a more competent catalytic machinery. Six model enzymes have been assayed in several model reactions in nonaqueous media. The results, rationalized in light of the presen…

Influence of hydrophobic matching on association of model transmembrane fragments containing a minimised glycophorin A dimerisation motif

AbstractThe principles that govern the folding and packing of membrane proteins are still not completely understood. In the present work, we have revisited the glycophorin A (GpA) dimerisation motif that mediates transmembrane (TM) helix association, one of the best-suited models of membrane protein oligomerisation. By using artificial polyleucine TM segments we have demonstrated in this study that a pattern of only five amino acids (GVxxGVxxT) promotes specific dimerisation. Further, we have used this minimised GpA motif to assess the influence of hydrophobic matching on the TM helix packing process in detergent micelles and found that this factor modulates helix–helix association and/or d…

Identification In Silico and In Vitro of Novel Trypanosomicidal Drug-Like Compounds

Atom-based bilinear indices and linear discriminant analysis are used to discover novel trypanosomicidal compounds. The obtained linear discriminant analysis-based quantitative structure–activity relationship models, using non-stochastic and stochastic indices, provide accuracies of 89.02% (85.11%) and 89.60% (88.30%) of the chemicals in the training (test) sets, respectively. Later, both models were applied to the virtual screening of 18 in-house synthesized compounds to find new pro-lead antitrypanosomal agents. The in vitro antitrypanosomal activity of this set against epimastigote forms of Trypanosoma cruzi is assayed. Predictions agree with experimental results to a great extent (16/18…

Core Histones Are Glutaminyl Substrates for Tissue Transglutaminase

Chicken erythrocyte core histones are glutaminyl substrates in the transglutaminase (TGase) reaction with monodansylcadaverine (DNC) as donor amine. The modification is very fast when compared with that of many native substrates of TGase. Out of the 18 glutamines of the four histones, nine (namely glutamine 95 of H2B; glutamines 5, 19, and 125 of H3; glutamines 27 and 93 of H4; and glutamines 24, 104, and 112 of H2A) are the amine acceptors in free histones. The use of Gln112 of H2A requires a temperature-dependent partial unfolding of the histone, showing that structural determinants are decisive for the glutamine specificity. The structures of H2A and H2B do not appreciably change upon mo…

A fluorescence spectroscopy study of the interaction of monocationic quinine with phospholipid vesicles Effect of the ionic strength and lipid composition

Abstract The interaction of monocationic quinine with zwitterionic dimyristoyl phosphatidylcholine (DMPC) and mixed negatively-charged dimyristoylphosphatidyl glycerol (DMPG) DMPC small unilamellar vesicles in the liquid-crystalline phase was investigated by steady-state fluorescence spectroscopy at pH 7 and 37°C. The maximum fluorescence emission peak at 383 nm, upon excitation at 335 nm, shifts to lower wavelength and decreases its intensity as the ratio between the total lipid and quinine concentrations increases. This indicates that in the membrane-bound state quinine is in an environment of low polarity, more deeply buried when anionic DMPG is present in the vesicle. For monoprotonated…

Macromolecules in ordered media: 7. Influence of ionic strength and bilayer composition on the association of polyelectrolytes to mixed liposomes

The effect of both the phospholipid composition of the bilayer and the ionic strength of the medium on the association between poly(2-vinyl pyridine) and mixed liposomes based on dimyristoyl phosphatidic acid and dimyristoyl phosphatidil choline was investigated using fluorescence spectroscopy. Intensity changes upon addition of vesicles have allowed us to estimate the extent of the association and data have been processed as association isotherms. We found that the association is enhanced by increasing both the ionic strength and the anionic phospholipid fraction. However, whereas the negative net charge of the bilayer strongly enhances the interaction, the presence of more and more electr…

Identification <i>In Silico</i> and <i>In Vitro</i> of Novel Trypanosomicidal Drug-like Compounds

Atom-based bilinear indices and linear discriminant analysis are used to discover novel trypanosomicidal compounds. The obtained linear discriminant analysis-based quantitative structure–activity relationship models, using non-stochastic and stochastic indices, provide accuracies of 89.02% (85.11%) and 89.60% (88.30%) of the chemicals in the training (test) sets, respectively. Later, both models were applied to the virtual screening of 18 in-house synthesized compounds to find new pro-lead antitrypanosomal agents. The in vitro antitrypanosomal activity of this set against epimastigote forms of Trypanosoma cruzi is assayed. Predictions agree with experimental results to a great extent (16/18…

<strong>Machine Learning and Atom-Based Quadratic Indices for Proteasome Inhibition Prediction </strong>

The atom-based quadratic indices are used in this work together with some machine learning techniques that includes: support vector machine, artificial neural network, random forest and k-nearest neighbor. This methodology is used for the development of two quantitative structure-activity relationship (QSAR) studies for the prediction of proteasome inhibition. A first set consisting of active and non-active classes was predicted with model performances above 85% and 80% in training and validation series, respectively. These results provided new approaches on proteasome inhibitor identification encouraged by virtual screenings procedures. .

Interaction of polyelectrolytes with oppositely charged micelles studied by fluorescence and liquid chromatography

It is studied by spectrofluorimetry the association of ionized cationic micelles (cetyltrimethylammonium bromide, CTAB) with oppositely charged polyelectrolyte [sodium poly(styrenesulfonate), PSSNa]. CTAB provokes a change in the fluorescence intensity emitted by PSSNa. The investigated surfactants form micelle-like aggregates before critical micellar concentration (CMC). Two approaches (binding and partition equilibrium) are used to obtain the association constant, KA, number of CTAB molecules in a binding site, N, and apparent partition coefficient, Γ. Analysis of the parameters as a function of polymer concentration and ionic strength μ is performed. The effect of μ shows an enhancement …

Polar/Ionizable Residues in Transmembrane Segments: Effects on Helix-Helix Packing

The vast majority of membrane proteins are anchored to biological membranes through hydrophobic alpha-helices. Sequence analysis of high-resolution membrane protein structures show that ionizable amino acid residues are present in transmembrane (TM) helices, often with a functional and/or structural role. Here, using as scaffold the hydrophobic TM domain of the model membrane protein glycophorin A (GpA), we address the consequences of replacing specific residues by ionizable amino acids on TM helix insertion and packing, both in detergent micelles and in biological membranes. Our findings demonstrate that ionizable residues are stably inserted in hydrophobic environments, and tolerated in t…

Membrane topology of gp41 and amyloid precursor protein: Interfering transmembrane interactions as potential targets for HIV and Alzheimer treatment

AbstractThe amyloid precursor protein (APP), that plays a critical role in the development of senile plaques in Alzheimer disease (AD), and the gp41 envelope protein of the human immunodeficiency virus (HIV), the causative agent of the acquired immunodeficiency syndrome (AIDS), are single-spanning type-1 transmembrane (TM) glycoproteins with the ability to form homo-oligomers. In this review we describe similarities, both in structural terms and sequence determinants of their TM and juxtamembrane regions. The TM domains are essential not only for anchoring the proteins in membranes but also have functional roles. Both TM segments contain GxxxG motifs that drive TM associations within the li…

Estimation of the Compatibility Between Poly(Methylmethacrylate) and Poly(Styrene Co Vinyl Phenol) Blends from Dilute Solution Measurements

Abstract The compatibility of poly(methyl methacrylate) (PMMA) with poly(styrene‐co‐vinyl phenol) (PS‐VPh) with two different contents of vinyl phenol, 5.8 and 7.2%, in dilute tetrahydrofuran solutions has been investigated by size exclusion chromatography and fluorescence spectroscopy at 25°C. The chromatographic technique permits the evaluation of the preferential solvation at different PMMA/PS‐VPh ratios. Changes in the fluorescence properties of PS‐VPh, caused by its association with PMMA, were used to obtain the fraction of copolymer bound to PMMA at diverse PMMA compositions. Both techniques agree quantitatively in every system, indicating that the association increases when the PMMA …

Miscibility of Blends of Biodegradable Polymers and Copolymers with Different Plasticizers

Miscibility between components of different plasticizer(A)/solvent(B)/biodegradable polymer(C) ternary systems with H-bonding has been compared. Systems were formed by two H-donor phenolic plasticizers, 4-nonylphenol (NP) and 4:4'-dihydroxydiphenylmethane (BPF); an H-acceptor solvent, epichlorohydrin (ECH); and H-acceptor poly(3-hydroxybutyrate) (PHB) or poly[(3-hydroxybutyrate)-co-(3-hydroxyvalerate)] (PHBHV) copolymers. Blend miscibility is the result of the balance of three competitive H-bondings: plasticizer self-association (AA), plasticizer-solvent (AB) and plasticizer-polymer (AC) interassociations. The strength and extent of such specific interactions have been experimentally monito…

Interactions of quinine with polyacrylic and poly-L-glutamic acids in aqueous solutions

Abstract Drug-loaded polymers provide an attractive form for controlled drug delivery systems. A proper knowledge of polymer–drug interactions can aid in the designing of polymers for various drug-delivery applications. In this paper we have investigated the interaction of a drug such as quinine, with synthetic macromolecules such as poly(acrylic acid), PAA, and poly(L-glutamic acid), PGA, at pH 7 and 37 °C by fluorescence spectroscopy and viscometry. The analysis of the binding isotherms revealed that the association process is positive cooperative up to a threshold concentration and then it is negative. In addition, the thermodynamic parameters vary along the isotherm. Results also sugges…

Long-chain fatty acyl-CoA esters induce lipase activation in the absence of a water-lipid interface.

In most lipases a mobile element or lid domain covers the catalytic site of the enzyme and the lid opening event, which usually proceed at a lipid-water interface, is required to form the catalytically competent lipase. We report here a noticeable increase in activity of two fungal lipases assayed in aqueous solution in absence of any interface when adding submicellar concentrations of amphipathic physiological molecules like long-chain acyl-CoAs. The catalytic activity was dramatically dependent on the acyl chain length of the amphiphile and could be related with a lid-opening process. Our data support that lipase activation can be triggered in the absence of a well-defined interface, and …

A semi-empirical approach for the simulation of circular dichroism spectra of gramicidin A in a model membrane

In an extension of our previous work (Bañó, M. C., Braco, L., and Abad, C. 1991. Biochemistry. 30:886-94), the kinetics of dissociation of gramicidin A double-stranded dimers into beta 6.3-helical monomers in small unilamellar vesicles prepared following different protocols, were investigated using in combination circular dichroism (CD) and high-performance liquid chromatography (HPLC). The analysis of the data from both techniques according to a two-component model strongly supports that any given CD pattern of gramicidin incorporated in the phospholipid bilayer can be deconvoluted essentially as a linear combination of the reference subspectra calculated for the double-stranded dimer and …

Determination of the relative quantum yields of the conformational species of autoassociating polypeptide gramicidin A in organic solvent using combined fluorescence spectroscopy and high performance liquid chromatography

Abstract A simple, novel method is proposed for the accurate determination of the relative quantum yields of each of the interconverting conformational species of the autoassociating polypeptide gramicidin A in organic solution. The method is based on fitting the experimental results obtained independently from fluorescence emission spectroscopy and high performance liquid chromatography. The fluorescence parameters obtained are discussed in terms of the structural features of the individual conformational species. The advantages of this approach and its possible application to other different organic solvents or to other autoassociating polypeptides are also considered.

A Study of Lipid-Lipid and Lipid-Polypeptide Interactions by High Performance Liquid Chromatography

Abstract Ternary systems containing phosphatidylcholine-cholesterol, phosphatidylcholine-gramicidin A or cholesterol-gramicidin A in tetrahydrofuran have been examined by high performance liquid chromatography. Preferential solvation of cholesterol and especially gramicidin A by phosphatidylcholine is observed. These results are interpreted in terms of hydrophobic interactions between membrane components.

Mark-Houwink Parameters of Biosynthetic Poly(γ-glutamic acid) in Aqueous Solution

A combined viscosity–light scattering–gel permeation chromatography (GPC) study was carried out on bacterially produced poly(γ-glutamic acid) (PGGA). PGGA samples with weight-average molecular weights ranging from 8×104 up to 8×105 g·mol–1 dissolved in phosphate buffer at 0.13 M ionic strength were used. It was found that the Mark–Houwink relation is acceptably obeyed, giving K and a values of 1.84×10–6 dL·g–1 and 1.16, respectively. As expected, GPC analysis showed that PGGA does not follow the universal calibration plot and that deviations can not be avoided by modifying the ionic strength.

Modeling Studies of the Phase Behavior of Monomer/Polymer/Disk Composites

The model developed by Balazs et al. to explain the phase behavior of polymer/clay composites is extended to monomer/polymer/clay composites, obtaining an expression for the free energy of a monomer/polymer/thin-disk mixture. By minimizing the free energy and calculating the chemical potentials of the three system components, phase diagrams for the monomer/disk and monomer/polymer/disk mixtures are contructed. Through the evolution and comparison of these diagrams, the effects of nanodisk size, polymer molecular mass and interaction parameters (temperature) on mixture stability and attained morphology are then studied.

Quantitative Prediction of Concentration Effects in Steric Exclusion Chromatography

Abstract A semiempirical model, based on a previous one quantitatively describing the dependence of the elution volume, V(cA), on the concentration of injected polymer, cA, in exclusion chromatography (SEC) at dilute solutions, has been developed. In the derived equation, concentration effects are mainly governed by the Huggins' coefficient, kA, and by the quadratic coefficient in the polynomial expansion of the reduced specific viscosity, kA. Because of the incertitudes on reliable kA and kA' values, these are respectively removed from the model through she Imai's equation and the empirical correlation kA' + 0.122=kA, here obtained. Thus, A predicted elution volumes besides polymer concent…

Conformational transitions of gramicidin A in phospholipid model membranes. A high-performance liquid chromatography assessment.

We have investigated the conformation of gramicidin A reconstituted in different phospholipid environments, small unilamellar vesicles, extensive bilayers, and micelles by exploiting a recently proposed experimental approach based on high-performance liquid chromatography [Bano et al. (1988) J. Chromatogr. 458, 105; Bano et al. (1989) FEBS Lett. 250, 67]. The method allows the separation of conformational species of the peptide namely, antiparallel double-stranded (APDS) dimers and β 6.3 -helical monomers, and quantitation of their proportions in the lipid environment. Various experimental parameters (e.g., nature of organic solvent, time of incubation in organic solvent, lipid-to-peptide m…

A Comparative Study of Nonlinear Machine Learning for the "In Silico" Depiction of Tyrosinase Inhibitory Activity from Molecular Structure.

In the preset report, for the first time, support vector machine (SVM), artificial neural network (ANN), Baye- sian networks (BNs), k-nearest neighbor (k-NN) are applied and compared on two "in-house" datasets to describe the tyrosinase inhibitory activity from the molecular structure. The data set Data I is used for the identification of tyrosi- nase inhibitors (TIs) including 701 active and 728 inactive compounds. Data II consists of active chemicals for potency estimation of TIs. The 2D TOMOCOMD-CARDD atom-based quadratic indices are used as molecular descriptors. The de- rived models show rather encouraging results with the areas under the Receiver Operating Characteristic (AURC) curve …

Characterization of Acylating and Deacylating Activities of an Extracellular Phospholipase A2 in a Water-Restricted Environment

The behavior of porcine pancreatic phospholipase A2 (ppPLA2) in monophasic low-water media has been explored, for the first time, in a systematic manner. It has been investigated how a number of variables can modulate both acylating and deacylating activities of the enzyme, and several interesting, unexpected results are presented. Among the most relevant, when placing ppPLA2 in the water-restricted environment, are the following: (i) it displays a remarkable alteration of its specificity toward the substrate polar head relative to all-water medium; (ii) it is quite severely inhibited by lysophosphatidylcholine (LPC), which has important implications, particularly concerning its acylation a…

Solution properties of polyelectrolytes. II. Comparison of the effects of both polyion and eluent salt concentrations on the calibrations in aqueous exclusion chromatography

The elution behaviour of charged water-soluble polymers (sodium polystyrene sulphonate) in both salt-free and NaCl-containing aqueous mobile phases has been investigated by size-exclusion chromatography. A comparative analysis has been established between the results as a function of polyion and salt concentrations, which demonstrates that the chromatographic behaviour is similar in both cases. The variation of the slopes of the different concentration-dependent calibration curves is discussed in terms of polyelectrolyte conformational changes. Finally, a semi-empirical model has been developed to predict the dependence of the elution volume in either polyion or salt concentration, and its …

Phosphatidylcholine-Gramicidin a Interaction Study in Non-Aqueous Solvent with a new HPLC-SEC Column

Abstract A new HPLC-SEC column, Ultrastyragel 500 A, has been assayed in order to check the separation between gramicidin A and phosphatidylcholine in tetrahydrofuran. The good resolution of both compounds has allowed the quantitation of lipid-polypeptide interaction in terms of the binding ratio parameter, BR. A BR value of 3.6 moles of phosphatidylcholine per mole of gramicidin A has been obtained for a 0.1 % (w/v) phosphatidylcholine solution as eluent. The suitability of this support for studies on lipid-polypeptide interactions in non-aqueous solvents is demonstrated.

Spectroscopic and chromatographic study of the interaction of nonionic micelles with the aluminum-morin complex

Abstract The interaction of the aluminum-morin system with different nonionic surfactants has been studied spectrophotometrically and by high-performance liquid chromatography. It has been found that ethylene oxide-propylene oxide condensate surfactants sensitize the spectrophotometric determination of aluminum with morin more than ethylene oxide condensate surfactants. Binding ratio parameters for the different surfactants with free morin have been also estimated from the analysis of chromatographic vacant peaks.

High-performance liquid chromatographic separation of modified and native melittin following transglutaminase-mediated derivatization with a dansyl fluorescent probe.

Abstract The 26-amino acid linear, amphiphilic peptide melittin was enzymatically modified with the fluorescent probe monodansylcadaverine using guinea pig liver transglutaminase and a fluorescent derivative of stoichiometry 1:1 was obtained. Reversed-phase and size-exclusion high-performance liquid chromatographic modes were tested in order to resolve the labelled peptide and native species. The influence of several operational variables was analysed and the elution conditions were optimized so that a satisfactory resolution could be achieved in both instances in a rapid, easy manner. Both chromatographic modes offer the possibility of accurate monitoring of the time course of the enzyme-m…

Binding of water-soluble, globular proteins to anionic model membranes

Abstract The role of electrostatics is studied in the adsorption of proteins to negatively charged (phosphatidylcholine/phosphatidylglycerol, PC/PG) and neutral (PC) small unilamellar vesicles (SUVs). For model proteins the interaction is monitored vs . pH at low ionic strength. The adsorption behaviour of lysozyme, myoglobin and albumin (isoelectronic point, p I 5–11) is investigated in SUVs, along with changes of the fluorescence emission spectra of the charged proteins, via their adsorption on SUVs. Significant adsorption of the proteins to negatively charged SUVs is found only at pH values, where the number of positive charge moieties exceeds the number of negative charge moieties on th…

A positional scanning combinatorial library of peptoids as a source of biological active molecules: identification of antimicrobials

9 pages, 4 figures, 2 schemes, 3 tables.-- PMID: 12959560 [PubMed].-- Printed version published in issue Sep-Oct 2003.-- Supporting information available at: http://pubs.acs.org/doi/suppl/10.1021/cc020075u

Study on resolution capacity and secondary mechanisms of three different TSK gels for organic SEC.

A comparative study on the elution behavior in size-exclusion chromatography (SEC) of three polymeric TSK gel packings, named H, H H R , and H X L types, is performed. The deviations of the universal calibration (u.c.) curves of seven solvent-polymer systems show evidence that the existence of secondary effects accompanying the main SEC mechanism. These secondary mechanisms are a consequence of the binary enthalpic interactions between the different components of the chromatographic system, such as polymer-solvent, polymer-gel and solvent-gel. However, in the present case, the observed deviations from a unique u.c. curve can mainly be attributed to adsorption of polymeric solutes (analytes)…

Solution properties of polyelectrolytes. I. Exclusion chromatography of sodium polystyrene sulphonate in salt-free water as eluent

The effect of concentration of sodium polystyrene sulphonate on the elution volume in exclusion chromatography has been investigated using salt-free water as eluent. A non-linear increase in the elution volume has been observed with increasing injected sample concentration. Concentration-dependent calibrations have been obtained which approach the universal calibration for uncharged polymers (dextrans) as polyelectrolyte concentration increases. The separation mechanisms are interpreted in terms of electrostatic and hydrodynamic interactions. A semi-empirical model is proposed to predict concentration effects of linear polyions in exclusion chromatography and good agreement has been found b…

Thermodynamic Study of Small Hydrophobic Ions at the Water–Lipid Interface

Abstract The thermodynamics of binding of two small hydrophobic ions such as norharman and tryptophan to neutral and negatively charged small unilamellar vesicles was investigated at pH 7.4 using fluorescence spectroscopy. Vesicles were formed at room temperature from dimyristoyl phosphatidylcholine (DMPC) or DMPC/dimyristoylphosphatidic acid and DMPC/dimyristoylphosphatidylglycerol. The changes in fluorescence properties were used to obtain association isotherms at variable membrane surface negative charge and at different ionic strengths. The binding of both ions was found to be quantitatively enhanced as the percentage of negative phospholipid increases in the membrane. Also, a decrease …

Bond-Based 2D Quadratic Fingerprints in QSAR Studies: Virtual and In vitro Tyrosinase Inhibitory Activity Elucidation

In this report, we show the results of quantitative structure–activity relationship (QSAR) studies of tyrosinase inhibitory activity, by using the bond-based quadratic indices as molecular descriptors (MDs) and linear discriminant analysis (LDA), to generate discriminant functions to predict the anti-tyrosinase activity. The best two models [Eqs (6) and (12)] out of the total 12 QSAR models developed here show accuracies of 93.51% and 91.21%, as well as high Matthews correlation coefficients (C) of 0.86 and 0.82, respectively, in the training set. The validation external series depicts values of 90.00% and 89.44% for these best two equations (6) and (12), respectively. Afterwards, a second …

HPLC demonstration that an all Trp--Phe replacement in gramicidin A results in a conformational rearrangement from beta-helical monomer to double-stranded dimer in model membranes.

We have taken advantage of our previously reported high performance liquid chromatographic (HPLC) strategy to investigate the conformational behavior of the optically reversed gramicidin M (gM-), an analog of gramicidin A with all tryptophans replaced by phenylalanines, in different model membranes. It is quantitatively demonstrated for the first time that once inserted in the lipid environment, gM- (unlike the native peptide) undergoes a conformational transition from beta-helical monomers to thermodynamically stable double-stranded dimers. This transition is faster the higher the incubation temperature and can be neatly observed in both small unilamellar phospholipid vesicles and lysophos…

Relative strength of H-bonding groups on biodegradable polymer-based blends in solution

The intermolecular hydrogen bonding interactions between poly(3-hydroxybutyrate) and poly(styrene-co-vinyl phenol) copolymers with mutual solvent epichlorohydrin were thoroughly investigated by steady-state fluorescence and viscosity techniques. Fluorescence spectroscopy along with viscosity technique was used to asses the intermolecular hydrogen bonding between poly-(3-hydroxybutyrate) and its blends with five copolymer samples of styrene–vinyl phenol, containing different proportions of vinyl phenol but similar average molecular weight and polydispersity index. In the case of very low OH contents (2–4 mol %), as expected, both components of poly(3-hydroxybutyrate) and poly(styrene-co-4-vi…