Activity coefficients of the electrolyte and the amino acid in water + NaNO(3) + glycine and water + NaCl + DL-methionine systems at 298.15 K

The activity coefficients at 298.15 K of glycine in water + NaNO(3) + glycine system and dl-methionine in water + NaCl + dl-methionine system are reported. The measurements were performed in an electrochemical cell with two ion selective electrodes, a cation and an anion ion selective electrode, each versus a double junction reference electrode. The concentrations of the electrolytes and the amino acids studied covered up to 1.0 molality electrolyte, 2.4 molality glycine and 0.2 molality dl-methionine. The results of the activity coefficients of glycine are compared with the activity coefficients of glycine in water + NaCl + glycine and water + KCl + glycine systems, obtained from the previous studies. The results show that the nature of both the cation and the anion of an electrolyte have significant effects on the activity coefficient of glycine in aqueous electrolyte solutions. The results also show that there are attractive interactions between the molecules of glycine and NaNO(3) and repulsive interactions between the molecules of dl-methionine and NaCl.     

Parametrization of direct and soft steric-undulatory forces between DNA double helical polyelectrolytes in solutions of several different anions and cations.

Directly measured forces between DNA helices in ordered arrays have been reduced to simple force coefficients and mathematical expressions for the interactions between pairs of molecules. The tabulated force parameters and mathematical expressions can be applied to parallel molecules or, by transformation, to skewed molecules of variable separation and mutual angle. This "toolbox" of intermolecular forces is intended for use in modelling molecular interactions, assembly, and conformation. The coefficients characterizing both the exponential hydration and the electrostatic interactions depend strongly on the univalent counterion species in solution, but are only weakly sensitive to anion type and temperature (from 5 to 50 degrees C). Interaction coefficients for the exponentially varying hydration force seen at spacings less than 10 to 15 A between surfaces are extracted directly from pressure versus interaxial distance curves. Electrostatic interactions are only observed at larger spacings and are always coupled with configurational fluctuation forces that result in observed exponential decay lengths that are twice the expected Debye-Huckel length. The extraction of electrostatic force parameters relies on a theoretical expression describing steric forces of molecules "colliding" through soft exponentially varying direct interactions.     

Effect of cation and anion of an electrolyte on apparent molar volume, isentropic compressibility and refractive index of glycine in aqueous solutions

Experiments at 298.15 K have been performed to measure the density, velocity of sound and refractive index in three water+glycine+electrolyte systems. The electrolytes studied were KCl, KNO3 and NaNO3. The values of apparent molar volume and isentropic compressibility of glycine in aqueous electrolyte solutions were calculated from the measured data. The results obtained in this study and those reported previously for water+glycine+NaCl system have been comparatively studied. The results show that the nature of both the cation and the anion of an electrolyte influence the behaviour of glycine in aqueous solutions. For all four electrolytes studied, the comparison shows a positive volume transfer for glycine from an electrolyte solution to a more concentrated solution of the same electrolyte. The results also show a negative apparent isentropic compressibility for glycine in the presence of the electrolytes studied. These effects indicate that the volume of a glycine molecule is larger in solutions with higher electrolyte concentration and the water molecules around the glycine molecules are less compressible than the water molecules in the bulk solution. These effects were attributed to the doubly charged behaviour of glycine and to the formation of physically bonded ion-pairs between the charged groups of glycine and the cation and the anion of the electrolyte.     

Hofmeister effects in supramolecular and biological systems

Specific ion effects, representative of near-universal Hofmeister phenomena, are illustrated in three different systems. These are the formation of supramolecular assemblies from cyclodextrins, the optical rotation of L-serine, and the growth rate of two kinds of microorganisms (Staphylococcus aureus and Pseudomonas aeruginosa). The strong specific ion effects can be correlated with the anion polarizabilities and related physico-chemical parameters. The results show the relevance of dispersion (non-electrostatic) forces in these phenomena.     

Activity coefficients of counterions and conformation in kappa-carrageenan systems

The optical rotation and the conductivity of kappa-carrageenans in aqueous solution have been investigated as functions of temperature in the presence of various electrolytes. The activity coefficients of sodium and potassium have been determined and correlated with the conformation. The potassium activity coefficient under ordered conformation is in agreement with a mechanism of dimerization.     

Staggered molecular packing in crystals of a collagen-like peptide with a single charged pair

The crystal structure of the triple-helical peptide, (Pro-Hyp-Gly)(4)-Glu-Lys-Gly-(Pro-Hyp-Gly)(5) has been determined to 1.75 A resolution. This peptide was designed to examine the effect of a pair of adjacent, oppositely charged residues on collagen triple-helical conformation and intermolecular interactions. The molecular conformation (a 7(5) triple helix) and hydrogen bonding schemes are similar to those previously reported for collagen triple helices and provides a second instance of water mediated N--H . . . O==C interchain hydrogen bonds for the amide group of the residue following Gly. Although stereochemically capable of forming intramolecular or intermolecular ion pairs, the lysine and glutamic acid side-chains instead display direct interactions with carbonyl groups and hydroxyproline hydroxyl groups or interactions mediated by water molecules. Solution studies on the EKG peptide indicate stabilization at neutral pH values, where both Glu and Lys are ionized, but suggest that this occurs because of the effects of ionization on the individual residues, rather than ion pair formation. The EKG structure suggests a molecular mechanism for such stabilization through indirect hydrogen bonding. The molecular packing in the crystal includes an axial stagger between molecules, reminiscent of that observed in D-periodic collagen fibrils. The presence of a Glu-Lys-Gly triplet in the middle of the sequence appears to mediate this staggered molecular packing through its indirect water-mediated interactions with backbone C==O groups and side chains.     

Protein-protein and protein-salt interactions in aqueous protein solutions containing concentrated electrolytes

Protein-protein and protein-salt interactions have been obtained for ovalbumin in solutions of ammonium sulfate and for lysozyme in solutions of ammonium sulfate, sodium chloride, potassium isothiocyanate, and potassium chloride. The two-body interactions between ovalbumin molecules in concentrated ammonium-sulfate solutions can be described by the DLVO potentials plus a potential that accounts for the decrease in free volume available to the protein due to the presence of the salt ions. The interaction between ovalbumin and ammonium sulfate is unfavorable, reflecting the kosmotropic nature of sulfate anions. Lysozyme-lysozyme interactions cannot be described by the above potentials because anion binding to lysozyme alters these interactions. Lysozyme-isothiocyanate complexes are strongly attractive due to electrostatic interactions resulting from bridging by the isothiocyanate ion. Lysozyme-lysozyme interactions in sulfate solutions are more repulsive than expected, possibly resulting from a larger excluded volume of a lysozyme-sulfate bound complex or perhaps, hydration forces between the lysozyme-sulfate complexes.     

Changes in the conformation of myo-inositol residues in triphosphoinositides]

Dependency of optical rotation of di- and triphosphoinositide water solutions on pH is investigated. The inversion of myo-inosite ring in triphosphoinositide molecule is found to take place at pH range corresponding to the transition of phosphomonoesters from monoanionic into dianionic form. Stabilization of optical rotation is observed at pH range, where one monoester phosphate group is in monoanionic form and the other--in dianionic form. Probably, triphosphoinositide is in the conformation (distorted boat) at this pH range. Diphosphoinositide does not change its optical rotation under pH changes of water or organic solutions. A contribution is estimated of free conformational energy into standard free energy of splitting triphosphoinositide phosphate group depending on pH value. It is concluded that low energy phosphate bond becomes high energy bond due to the free electrostatic interaction of dianionic phosphate group with other negatively charged group in sin-clynal conformation.     

Evidence for the Interaction of Nucleotides with Immobilized Amino-acids and its Significance for the Origin of the Genetic Code

ONE of the essential relationships between nucleic acids and amino-acids in present biological systems and perhaps in evolutionary precursors to these systems is expressed in binding and recognition interactions. Such interactions depend on the size, composition and conformation of the interacting species^1–8. When the two reacting species are simple (that is, when neither is polymeric) one cannot expect to observe “specificity” of the sort implied in the biological use of the term. Working with monomeric species in aqueous media permits the effects of individual factors to be assessed so that more complex interactions between these molecules can be understood and their evolutionary potential evaluated.     

VALENCY RULE AND ALLEGED HOFMEISTER SERIES IN THE COLLOIDAL BEHAVIOR OF PROTEINS : II. THE INFLUENCE OF SALTS.

It is shown by the older experiments by Loeb and by the experiments reported in this paper that the effect of salts on the membrane potentials, osmotic pressure, swelling of gelatin chloride, and that type of viscosity which is due to the swelling of protein particles, depends only on the valency but not on the chemical nature of the anion of the salt, and that the cation of the salt has no effect on these properties, if the pH of the protein solution or protein gel is not altered by the salt. The so called Hofmeister series of salt effects on these four properties are purely fictitious and due to the failure of the former authors to measure the hydrogen ion concentration of their protein solutions or gels and to compare the effects of salts at the same pH of the protein solution or the protein gel. These results confirm the older experiments of Loeb and together they furnish a further proof for the correctness of the idea that the influence of electrolytes on the four properties of proteins is determined by membrane equilibria. Such properties of proteins which do not depend on membrane equilibria, such as solubility or cohesion, may be affected not only by the valency but also by the chemical nature of the ions of a salt.     

INFLUENCE OF A SLIGHT MODIFICATION OF THE COLLODION MEMBRANE ON THE SIGN OF THE ELECTRIFICATION OF WATER

1. It is shown that collodion membranes which have received one treatment with a 1 per cent gelatin solution show for a long time (if not permanently) afterwards a different osmotic behavior from collodion membranes not treated with gelatin. This difference shows itself only towards solutions of those electrolytes which have a tendency to induce a negative electrification of the water particles diffusing through the membrane, namely solutions of acids, acid salts, and of salts with trivalent and tetravalent cations; while the osmotic behavior of the two types of membranes towards solutions of salts and alkalies, which induce a positive electrification of the water particles diffusing through the membrane, is the same. 2. When we separate solutions of salts with trivalent cation, e.g. LaCl₃ or AlCl₃, from pure water by a collodion membrane treated with gelatin, water diffuses rapidly into the solution; while no water diffuses into the solution when the collodion membrane has received no gelatin treatment. 3. When we separate solutions of acid from pure water by a membrane previously treated with gelatin, negative osmosis occurs; i.e., practically no water can diffuse into the solution, while the molecules of solution and some water diffuse out. When we separate solutions of acid from pure water by collodion membranes not treated with gelatin, positive osmosis will occur; i.e., water will diffuse rapidly into the solution and the more rapidly the higher the valency of the anion. 4. These differences occur only in that range of concentrations of electrolytes inside of which the forces determining the rate of diffusion of water through the membrane are predominantly electrical; i.e., in concentrations from 0 to about M/16. For higher concentrations of the same electrolytes, where the forces determining the rate of diffusion are molecular, the osmotic behavior of the two types of membranes is essentially the same. 5. The differences in the osmotic behavior of the two types of membranes are not due to differences in the permeability of the membranes for solutes since it is shown that acids diffuse with the same rate through both kinds of membranes. 6. It is shown that the differences in the osmotic behavior of the two types of collodion membranes towards solutions of acids and of salts with trivalent cation are due to the fact that in the presence of these electrolytes water diffuses in the form of negatively charged particles through the membranes previously treated with gelatin, and in the form of positively charged particles through collodion membranes not treated with gelatin. 7. A treatment of the collodion membranes with casein, egg albumin, blood albumin, or edestin affects the behavior of the membrane towards salts with trivalent or tetravalent cations and towards acids in the same way as does a treatment with gelatin; while a treatment of the membranes with peptone prepared from egg albumin, with alanine, or with starch has no such effect.     

THE INFLUENCE OF ELECTROLYTES ON THE ELECTRIFICATION AND THE RATE OF DIFFUSION OF WATER THROUGH COLLODION MEMBRANES

1. When pure water is separated by a collodion membrane from a watery solution of an electrolyte the rate of diffusion of water is influenced not only by the forces of gas pressure but also by electrical forces. 2. Water is in this case attracted by the solute as if the molecules of water were charged electrically, the sign of the charge of the water particles as well as the strength of the attractive force finding expression in the following two rules, (a) Solutions of neutral salts possessing a univalent or bivalent cation influence the rate of diffusion of water through a collodion membrane, as if the water particles were charged positively and were attracted by the anion and repelled by the cation of the electrolyte; the attractive and repulsive action increasing with the number of charges of the ion and diminishing inversely with a quantity which we will designate arbitrarily as the "radius" of the ion. The same rule applies to solutions of alkalies. (b) Solutions of neutral or acid salts possessing a trivalent or tetravalent cation influence the rate of diffusion of water through a collodion membrane as if the particles of water were charged negatively and were attracted by the cation and repelled by the anion of the electrolyte. Solutions of acids obey the same rule, the high electrostatic effect of the hydrogen ion being probably due to its small "ionic radius." 3. The correctness of the assumption made in these rules concerning the sign of the charge of the water particles is proved by experiments on electrical osmose. 4. A method is given by which the strength of the attractive electric force of electrolytes on the molecules of water can be roughly estimated and the results of these measurements are in agreement with the two rules. 5. The electric attraction of water caused by the electrolyte increases with an increase in the concentration of the electrolyte, but at low concentrations more rapidly than at high concentrations. A tentative explanation for this phenomenon is offered. 6. The rate of diffusion of an electrolyte from a solution to pure solvent through a collodion membrane seems to obey largely the kinetic theory inasmuch as the number of molecules of solute diffusing through the unit of area of the membrane in unit time is (as long as the concentration is not too low) approximately proportional to the concentration of the electrolyte and is the same for the same concentrations of LiCl, NaCl, MgCl₂, and CaCl₂.     

Effect of temperature on the conformation of native DNA in aqueous solutions of various electrolytes

Effect of the temperature on the conformation of the native DNA molecule in solution of different electrolytes (LiCl, NaCl, KCl, CsCl, Gu-HCl) at ionic strengths mu = 5; 0.1; 0.01; 0.005 and temperatures ranging from 10 to 40 degrees C were studied by the methods of flow birefringence and viscometry. The experiments showed that the value of intrinsic viscosity [eta] of DNA increases at increase of temperatures in solutions of all the chlorides studied, excluding guanidine. The effect of temperature on the value of [eta] doesn't depend on the type of the cation at a fixed value of mu and is elevated when mu decreases. The observed alterations of the value of [eta] for DNA in water-salt solutions at different temperatures can be explained by an increase in the hydration of the alkaline ions at temperature increase. The experiments showed the specificity of the effect of different ions on the dimensions of the DNA molecule in solution. The data on optical anisotropy of the DNA molecule testify, that the thermodynamic rigidity of the latter doesn't depend on the temperature of solutions of different electrolytes in the temperature range studied.     

Amylose conformational transitions in binary DMSO/water mixtures

The effects on amylose conformation of percentage water in dimethyl sulfoxide (DMSO)/water mixtures were measured by following changes in specific optical rotation, limiting viscosity number, and ¹³C-NMR chemical shifts. The temperature dependence of specific optical rotation showed differences in amylose conformation at four chosen ratios of dimethyl sulfoxide/water. An amylose conformational change was also deduced from ¹³C-NMR chemical shift data. Changes in limiting viscosity of amylose in different proportions of DMSO/water, and the effect of tetramethylurea on the specific rotation of amylose, indicate that intramolecular hydrogen bonding decreases with increased water content. 66.6% DMSO appears to be a crossover concentration, below which the helical conformation is progressively lost as water is added. When water content is over 60%, transition to a conformation which allows iodine complexation to take place is complete. A transition of amylose conformation from helix to loose helix to random coil with increasing water content was deduced from the experimental results.     

1-anilino-8-naphthalene sulfonate as a protein conformational tightening agent

1-Anilino-8-naphthalene sulfonate (ANS) anion is conventionally considered to bind to preexisting hydrophobic (nonpolar) surfaces of proteins, primarily through its nonpolar anilino-naphthalene group. Such binding is followed by an increase in ANS fluorescence intensity, similar to that occurring when ANS is dissolved in organic solvents. It is generally assumed that neither the negative sulfonate charge on the ANS, nor charges on the protein, participate significantly in ANS-protein interaction. However, titration calorimetry has demonstrated that most ANS binding to a number of proteins occurs through electrostatic forces, in which ion pairs are formed between ANS sulfonate groups and cationic groups on the proteins (D. Matulis and R. E. Lovrien, Biophys. J., 1998, Vol. 74, pp. 1-8). Here we show by viscometry and diffusion coefficient measurements that bovine serum albumin and gamma-globulin, starting from their acid-expanded, most hydrated conformations, undergo extensive molecular compaction upon ANS binding. As the cationic protein binds negatively charged ANS anion it also takes up positively charged protons from water to compensate the effect of the negative charge, and leaves the free hydroxide anions in solution thus shifting pH upward (the Scatchard-Black effect). These results indicate that ANS is not always a definitive reporter of protein molecular conformation that existed before ANS binding. Instead, ANS reports on a conformationally tightened state produced by the interplay of ionic and hydrophobic characters of both protein and ligand.     

Recognition schemes for protein-nucleic acid interactions

The molecular forces involved in protein-nucleic acid interaction are electrostatic, stacking and hydrogen-bonding. These interactions have a certain amount of specificity due to the directional nature of such interactions and the spatial contributions of the steric effects of different substituent groups. Quantum chemical calculations on these interactions have been reported which clearly bring out such features. While the binding energies for electrostatic interactions are an order of magnitude higher, the differences in interaction energies for structures stabilised by hydrogen-bonding and stacking are relatively small. Thus, the molecular interactions alone cannot explain the highly specific nature of binding observed in certain segments of proteins and nucleic acids. It is therefore logical to assume that the sequence dependent three dimensional structures of these molecules help to place the functional groups in the correct geometry for a favourable interaction between the two molecules. We have carried out 2D-FT nuclear magnetic resonance studies on the oligonucleotide d-GGATCCGGATCC. This oligonucleotide sequence has two binding sites for the restriction enzyme Bam H1. Our studies indicate that the conformation of this DNA fragment is predominantly B-type except near the binding sites where the ribose ring prefers a³E conformation. This interesting finding raises the general question about the presence of specificity in the inherent backbone structures of proteins and nucleic acids as opposed to specific intermolecular interactions which may induce conformational changes to facilitate such binding.     

Water absorbency by wool fibers: Hofmeister effect

Wool is a complex material, composed of cuticle and epicuticle cells, surrounded by a cell membrane complex. Wool fibers absorb moisture from air, and, once immersed in water, they take up considerable amounts of liquid. The water absorbency parameter can be determined from weight gain, according to a standard method, and used to quantify this phenomenon. In this paper we report a study on the water absorbency (or retention) of untreated wool fibers in the presence of aqueous 1 M salt solutions at 29 degrees C and a relative humidity of either 33% or 56%. The effect of anions was determined by selecting a wide range of different sodium salts, while the effect of cations was checked through some chlorides and nitrates. Our results show a significant specific ion and ion pair "Hofmeister" effects, that change the amount of water absorbed by the fibers. To understand this phenomenon, the water absorbency parameter (A(w)) is compared to different physicochemical parameters such as the lyotropic number, free energy of hydration of ions, molar surface tension increment, polarizability, refractive index increment, and molar refractivity. The data indicate that this Hofmeister phenomenon is controlled by dispersion forces that depend on the polarizability of ionic species, their adsorption frequencies, the solvent, and the substrate. These dispersion forces dominate the behavior in concentrated solutions. They are in accord with new developing theories of solutions and molecular interactions in colloidal systems that account for Hofmeister effects.     

Force generation in single conventional actomyosin complexes under high dynamic load

The mechanical load borne by a molecular motor affects its force, sliding distance, and its rate of energy transduction. The control of ATPase activity by the mechanical load on a muscle tunes its efficiency to the immediate task, increasing ATP hydrolysis as the power output increases at forces less than isometric (the Fenn effect) and suppressing ATP hydrolysis when the force is greater than isometric. In this work, we used a novel 'isometric' optical clamp to study the mechanics of myosin II molecules to detect the reaction steps that depend on the dynamic properties of the load. An actin filament suspended between two beads and held in separate optical traps is brought close to a surface that is sparsely coated with motor proteins on pedestals of silica beads. A feedback system increases the effective stiffness of the actin by clamping the force on one of the beads and moving the other bead electrooptically. Forces measured during actomyosin interactions are increased at higher effective stiffness. The results indicate that single myosin molecules transduce energy nearly as efficiently as whole muscle and that the mechanical control of the ATP hydrolysis rate is in part exerted by reversal of the force-generating actomyosin transition under high load without net utilization of ATP.     

Conformation and structure of mildly heat-treated ovalbumin in dilute solutions and gel formation at higher protein concentrations

The conformation of heat-denatured ovalbumin aggregates has been examined at several concentrations and pH values, using measurements of optical rotation dispersion (ORD), circular dichroism (CD) and viscosity. The protein was subjected to heating at relatively low temperatures, ranging from 48.5 to 76 degrees; the particular temperature chosen depended on pH. The heat treatment was sufficient to remove the ability of the molecules to absorb heat on re-heating. The denatured molecules were shown to be rather compact, i.e. not much larger than the native molecule, and to retain a significant amount of secondary structure; this was also the case for molecules present in small aggregates. It is suggested that this type of ovalbumin monomer builds three-dimensional networks in denatured solutions at higher concentrations, and that gelation should be looked upon as arising from surface contacts between hydrated globules. The present results also imply that such globules have gelation properties which depend on whether pH is acidic or basic compared to the isoelectric point of the protein.