Enthalpic Contributions Research Articles

Polyelectrolyte Complexation When Considering the Counterion-Mediated Hydrogen Bonding.

In this work, we have investigated a pH-modulated complexation between two oppositely charged strong polyelectrolytes to demonstrate the effect of counterion-mediated hydrogen bonding (CMHB) on polyelectrolyte complexation. We have found that such a pH-modulated complexation cannot be understood without considering the CMHB. Thermodynamically, the effect of CMHB on the polyelectrolyte complexation is manifested by the alteration of both enthalpic and entropic contributions to the free energy change. The pH-dependent intrinsic ion-pairing and complex coacervation processes of the polyelectrolyte complexation can be understood when considering the CMHB. Our study demonstrates that both the extent of polyelectrolyte complex formation in bulk solutions and the formation of polyelectrolyte multilayers on surfaces are controlled by the pH-dependent intrinsic ion-pairing process. Furthermore, on the basis of the pH-dependent intrinsic ion pairing, the properties of the multilayers can be tuned by pH. This work provides a new strategy to control the polyelectrolyte complexation with counterions and will inspire new ideas for building advanced polyelectrolyte materials.

Kinetic Modeling, Thermodynamic Approach and Molecular Dynamics Simulation of Thermal Inactivation of Lipases from Burkholderia cepacia and Rhizomucor miehei.

The behavior against temperature and thermal stability of enzymes is a topic of importance for industrial biocatalysis. This study focuses on the kinetics and thermodynamics of the thermal inactivation of Lipase PS from B. cepacia and Palatase from R. miehei. Thermal inactivation was investigated using eight inactivation models at a temperature range of 40–70 °C. Kinetic modeling showed that the first-order model and Weibull distribution were the best equations to describe the residual activity of Lipase PS and Palatase, respectively. The results obtained from the kinetic parameters, decimal reduction time (D and tR), and temperature required (z and z’) indicated a higher thermal stability of Lipase PS compared to Palatase. The activation energy values (Ea) also indicated that higher energy was required to denature bacterial (34.8 kJ mol−1) than fungal (23.3 kJ mol−1) lipase. The thermodynamic inactivation parameters, Gibbs free energy (ΔG#), entropy (ΔS#), and enthalpy (ΔH#) were also determined. The results showed a ΔG# for Palatase (86.0–92.1 kJ mol−1) lower than for Lipase PS (98.6–104.9 kJ mol−1), and a negative entropic and positive enthalpic contribution for both lipases. A comparative molecular dynamics simulation and structural analysis at 40 °C and 70 °C were also performed.

Dynamics and disorder: on the stability of pyrazinamide polymorphs.

This article focuses on the structure and relative stability of four pyrazinamide polymorphs. New single crystal X-ray diffraction data collected for all forms at 10 K and 122 K are presented. By combining periodic ab initio DFT calculations with normal-mode refinement against X-ray diffraction data, both enthalpic and entropic contributions to the free energy of all polymorphs are calculated. On the basis of the estimated free energies, the stability order of the polymorphs as a function of temperature and the corresponding solid state phase transition temperatures are anticipated. It can be concluded that the α and γ forms have higher vibrational entropy than that of the β and δ forms and therefore they are significantly more stabilized at higher temperatures. Due to the entropy which arises from the disorder in γ form, it overcomes form α and is the most stable form at temperatures above ∼500 K. Our findings are in qualitative agreement with the experimental calorimetry results.

Structure, dynamics, and molecular inhibition of the Staphylococcus aureus m1A22-tRNA methyltransferase TrmK

The enzyme m1A22-tRNA methyltransferase (TrmK) catalyzes the transfer of a methyl group to the N1 of adenine 22 in bacterial tRNAs. TrmK is essential for Staphylococcus aureus survival during infection but has no homolog in mammals, making it a promising target for antibiotic development. Here, we characterize the structure and function of S. aureus TrmK (SaTrmK) using X-ray crystallography, binding assays, and molecular dynamics simulations. We report crystal structures for the SaTrmK apoenzyme as well as in complexes with methyl donor SAM and co-product product SAH. Isothermal titration calorimetry showed that SAM binds to the enzyme with favorable but modest enthalpic and entropic contributions, whereas SAH binding leads to an entropic penalty compensated for by a large favorable enthalpic contribution. Molecular dynamics simulations point to specific motions of the C-terminal domain being altered by SAM binding, which might have implications for tRNA recruitment. In addition, activity assays for SaTrmK-catalyzed methylation of A22 mutants of tRNALeu demonstrate that the adenine at position 22 is absolutely essential. In silico screening of compounds suggested the multifunctional organic toxin plumbagin as a potential inhibitor of TrmK, which was confirmed by activity measurements. Furthermore, LC-MS data indicated the protein was covalently modified by one equivalent of the inhibitor, and proteolytic digestion coupled with LC-MS identified Cys92 in the vicinity of the SAM-binding site as the sole residue modified. These results identify a cryptic binding pocket of SaTrmK, laying a foundation for future structure-based drug discovery.

The Thermodynamic Stability of Membrane Proteins in Micelles and Lipid Bilayers Investigated with the Ferrichrom Receptor FhuA

Extraction of integral membrane proteins into detergents for structural and functional studies often leads to a strong loss in protein stability. The impact of the lipid bilayer on the thermodynamic stability of an integral membrane protein in comparison to its solubilized form in detergent was examined and compared for FhuA from Escherichia coli and for a mutant, FhuAΔ5-160, lacking the N-terminal cork domain. Urea-induced unfolding was monitored by fluorescence spectroscopy to determine the effective free energies Delta G{^text{o}_{rm u}} of unfolding. To obtain enthalpic and entropic contributions of unfolding of FhuA, Delta G{^text{o}_{rm u}} were determined at various temperatures. When solubilized in LDAO detergent, wt-FhuA and FhuAΔ5-160 unfolded in a single step. The 155-residue cork domain stabilized wt-FhuA by DeltaDelta G{^text{o}_{rm u}} ~ 40 kJ/mol. Reconstituted into lipid bilayers, wt-FhuA unfolded in two steps, while FhuAΔ5-160 unfolded in a single step, indicating an uncoupled unfolding of the cork domain. For FhuAΔ5-160 at 35 °C, Delta G{^text{o}_{rm u}} increased from ~ 5 kJ/mol in LDAO micelles to about ~ 20 kJ/mol in lipid bilayers, while the temperature of unfolding increased from TM ~ 49 °C in LDAO micelles to TM ~ 75 °C in lipid bilayers. Enthalpies Delta H{_{rm M}^text{o}}were much larger than free energies Delta G{^text{o}_{rm u}} , for FhuAΔ5-160 and for wt-FhuA, and compensated by a large gain of entropy upon unfolding. The gain in conformational entropy is expected to be similar for unfolding of FhuA from micelles or bilayers. The strongly increased TM and Delta H{_{rm M}^text{o}} observed for the lipid bilayer-reconstituted FhuA in comparison to the LDAO-solubilized forms, therefore, very likely arise from a much-increased solvation entropy of FhuA in bilayers.Graphical abstract

Enthalpic and entropic contributions to interleaflet coupling drive domain registration and antiregistration in biological membrane.

Biological membrane is a complex self-assembly of lipids, sterols, and proteins organized as a fluid bilayer of two closely stacked lipid leaflets. Differential molecular interactions among its diverse constituents give rise to heterogeneities in the membrane lateral organization. Under certain conditions, heterogeneities in the two leaflets can be spatially synchronized and exist as registered domains across the bilayer. Several contrasting theories behind mechanisms that induce registration of nanoscale domains have been suggested. Following a recent study showing the effect of position of lipid tail unsaturation on domain registration behavior, we decided to develop an analytical theory to elucidate the driving forces that create and maintain domain registry across leaflets. Towards this, we formulated a Hamiltonian for a stacked lattice system where site variables capture the lipid molecular properties such as the position of unsaturation and various other interactions that could drive phase separation and interleaflet coupling. We solve the Hamiltonian using Monte Carlo simulations and create a complete phase diagram that reports the presence or absence of registered domains as a function of various Hamiltonian parameters. We find that the interleaflet coupling should be described as a competing enthalpic contribution due to interaction of lipid tail termini, primarily due to saturated-saturated interactions, and an interleaflet entropic contribution from overlap of unsaturated tail termini. A higher position of unsaturation is seen to provide weaker interleaflet coupling. Thermodynamically stable nanodomains could also be observed for certain points in the parameter space in our bilayer model, which were further verified by carrying out extended Monte Carlo simulations. These persistent noncoalescing registered nanodomains close to the lower end of the accepted nanodomain size range also point towards a possible "nanoscale" emulsion description of lateral heterogeneities in biological membrane leaflets.

Design of molecularly imprinted polymer materials relying on hydrophobic interactions

The design of molecularly imprinted polymers (MIPs) for the specific binding of hydrophobic molecules (mitotane) was investigated by considering monomers and cross-linking agents with variable hydrophobic character, methyl methacrylate, butyl methacrylate and lauryl methacrylate as functional monomers, and ethylene glycol dimethacrylate and 1,6-hexanediol dimethacrylate as cross-linking agents. MIPs were bound to a porous silica support by means of a radical transfer reaction with grafted 3-mercaptopropyl groups. Equilibrium adsorption isotherms to molecularly imprinted and non-imprinted materials were modeled with the Volmer and Langmuir–Volmer isotherms, giving the thermodynamic parameters of adsorption. The most hydrophobic monomer provided the highest selective adsorption although strong non-selective adsorption and the intrinsic softness of poly(lauryl methacrylate) is detrimental to selectivity. Adsorption was exothermic with a predominant enthalpic contribution. Adsorption kinetics were fast due to the high accessibility of molecular imprints on the surface of the thin MIP layer coating the porous silica support.

Negatively charged poly(N-isopropyl acrylamide-co-methacrylic acid)/polyacrylamide semi-IPN hydrogels: Correlation between swelling and compressive elasticity

A series of thermoresponsive semi-interpenetrating polymer network (semi-IPN) based on linear polyacrylamide (PAAm), N-isopropylacrylamide (NIPA) and methacrylic acid (MA) were synthesized by varying linear polymer content via free radical polymerization. An interpenetrated network of P(NIPA-MA)/PAAm was designed to modulate elasticity by varying inner composition and to improve rate of swelling-deswelling phenomenon. Correlation between swelling and compression elasticity was demonstrated. Inclusion of linear PAAm chains with high molecular weight (Mw = 9.639 × 105 g/mol) into P(NIPA-MA) network induced physical entanglements, increased apparent crosslinking density νeand enlarged compressive elasticity. Dependence of νeon linear polymer content was expressed as a cubic polynomial function. Extent of swelling of semi-IPN P(NIPA-MA)/PAAm gels was sensitive to presence of linear PAAm chains. Existence of linear polymer decreased apparent ionic group density and increased crosslinking density compared to that of copolymer P(NIPA-MA) network which in turn decreased equilibrium swelling. A significant increase in swelling/shrinking rate was observed in the presence of linear PAAm. Due to ionization of carboxylic acid groups in P(NIPA-MA) network, semi-IPN P(NIPA-MA)/PAAm gels showed different degrees of swelling depending on linear PAAm content and temperature of swelling medium. Semi-IPNs exhibited phase transition temperatures shifted higher temperature, suggesting physical entanglements between P(NIPA-MA) network and linear PAAm. Increasing swelling temperature resulted in an increase in Flory-Huggins interaction parameter. Entropic contributionχSincreased and enthalpic contribution χH decreased with PAAm content. The results showed that the prepared semi-IPNs are a suitable adsorbent for adsorption of the cationic dye Methyl violet (MV). Adsorption kinetics followed a pseudo-second-order model and exhibited a two-stage intra-particle diffusion. A detailed understanding of physicochemical properties of negatively charged thermoresponsive semi-IPNs in relation to structural architecture is an important criterion in many biomedical and pharmaceutical applications.

A β-NMR study of the depth, temperature, and molecular-weight dependence of secondary dynamics in polystyrene: Entropy-enthalpy compensation and dynamic gradients near the free surface.

We investigated the depth, temperature, and molecular-weight (MW) dependence of the γ-relaxation in polystyrene glasses using implanted 8Li+ and β-detected nuclear magnetic resonance. Measurements were performed on thin films with MW ranging from 1.1 to 641 kg/mol. The temperature dependence of the average 8Li spin-lattice relaxation time (T1 avg) was measured near the free surface and in the bulk. Spin-lattice relaxation is caused by phenyl ring flips, which involve transitions between local minima over free-energy barriers with enthalpic and entropic contributions. We used transition state theory to model the temperature dependence of the γ-relaxation, and hence T1 avg. There is no clear correlation of the average entropy of activation (Δ‡S̄) and enthalpy of activation (Δ‡H̄) with MW, but there is a clear correlation between Δ‡S̄ and Δ‡H̄, i.e., entropy-enthalpy compensation. This results in the average Gibbs energy of activation, Δ‡Ḡ, being approximately independent of MW. Measurements of the temperature dependence of T1 avg as a function of depth below the free surface indicate the inherent entropic barrier, i.e., the entropy of activation corresponding to Δ‡H̄ = 0, has an exponential dependence on the distance from the free surface before reaching the bulk value. This results in Δ‡Ḡ near the free surface being lower than the bulk. Combining these observations results in a model where the average fluctuation rate of the γ-relaxation has a "double-exponential" depth dependence. This model can explain the depth dependence of 1/T1 avg in polystyrene films. The characteristic length of enhanced dynamics is ∼6nm and approximately independent of MW near room temperature.

Micellization and thermodynamics study of n-alkyl-4-methylpyridinium bromides in water and mixed water–ethanol media

The micellization behaviors of n-decyl-4-methylpyridinium bromide, n-dodecyl-4-methylpyridinium bromide, and n-tetradecyl-4-methylpyridinium bromide were studied in water and mixed water–ethanol media by using conductivity measurements over the temperature range of 293.15–318.15 K. The critical micelle concentrations (CMC) of all n-alkyl-4-methyl pyridinium bromides studied increased with increasing temperature, but they decreased with increasing hydrocarbon chain length. Their CMC values of n-decyl-4-methylpyridinium bromide, n-dodecyl-4-methylpyridinium bromide, and n-tetradecyl-4-methylpyridinium bromide in water at the temperature range of 293.15–318.15 K were determined as in the range of 43.62–46.42, 10.45–11.57, and 2.78–3.20 mM, respectively. Moreover, the lowest CMC values of n-decyl-4-methylpyridinium bromide, n-dodecyl-4-methylpyridinium bromide, and n-tetradecyl-4-methylpyridinium bromide were found with 15, 5, and 5 vol% ethanol additions, respectively. The calculated negative ΔGm0 and ΔHm0 values proved that their micellizations in water and mixed water–ethanol media were spontaneous and exothermic. Also, ΔGm0 values became the more negative with the longer the hydrocarbon chain length. Furthermore, their micellization processes in water were determined to be entropy controlled, however enthalpic contributions in the mixed water–ethanol systems started to predominate with increasing ethanol concentrations and/or temperature.

Mechanical Size Effect of Freestanding Nanoconfined Polymer Films

While elastic properties of nanoconfined polymer films have been recognized to show departures from bulk behavior, a careful understanding of the origins of mechanical size effects remains weak. Here, we report a significant mechanical stiffening of freestanding ultrathin poly(methyl methacrylate) films of varying thicknesses (6–200 nm) through atomic force microscopy deflection measurements at ambient conditions. After excluding the substrate influence, the stiffening mechanism is linked to extended chain conformations based on small-angle X-ray scattering and infrared nanoscopic characterization. We advocate that the entropic elasticity of individual chains plays a significant role in polymer mechanics in nanoscale thickness films, where the entanglement density is apparently low, with chains oriented in the plane of the film, unlike a bulk polymer. Molecular dynamics simulations further unveil the dominance of entropic contributions over enthalpic contributions to the chain stiffness that endows polymer films with higher load-bearing capacity and accounts for the stiffening at the nanoscale. The results presented herein provide a mechanistic understanding of molecular origins of the size effect, serving as a potent design strategy for accessing high-performance polymer-based devices.

To Be or Not To Be? that is the Entropic, Enthalpic, and Molecular Interaction Dilemma in the Formation of (Water)20 Clusters and Methane Clathrate.

A detailed analysis under a comprehensive set of theoretical and computational tools of the thermodynamical factors and of the intermolecular interactions behind the stabilization of a well known set of (water)20 cavities and of the methane clathrate is offered in this work. Beyond the available reports of experimental characterization at extreme conditions of most of the systems studied here, all clusters should be amenable to experimental detection at 1 atm and moderate temperatures since 280 K marks the boundary at which, ignoring reaction paths, formation of all clusters is no longer spontaneous from the 20H2 O→(H2 O)20 and CH4 +20H2 O→CH4 @512 processes. As a function of temperature, a complex interplay leading to the free energy of formation occurs between the destabilizing entropic contributions, mostly due to cluster vibrations, and the stabilizing enthalpic contributions, due to intermolecular interactions and the PV term, is best illustrated by the highly symmetric 512 cage consistently showing signs of stronger intermolecular bonding despite having smaller binding energy than the other clusters. A fluxional wall of attractive non-covalent interactions, arising because of the cumulative effect of a large number of tiny individual charge transfers to the interstitial region, plays a pivotal role stabilizing the CH4 @512 clathrate.

Augmented Self-Association by Electrostatic Forces in Thienopyrrole-Fused Thiadiazoles that Contain an Ester instead of an Ether Linker.

During the self-assembly of π-conjugated molecules, linkers and substituents can potentially add supportive noncovalent intermolecular interactions to π-stacking interactions. Here, we report the self-assembly behavior of thienopyrrole-fused thiadiazole (TPT) fluorescent dyes that possess ester or ether linkers and dodecyloxy side chains in solution and the condensed phase. A comparison of the self-association behavior of the ester- and ether-bridged compounds in solution using detailed UV-vis, fluorescence, and NMR spectroscopic studies revealed that the subtle replacement of the ether linkers by ester linkers leads to a distinct increase in the association constant (ca. 3-4 fold) and the enthalpic contribution (ca. 3 kcal mol-1 ). Theoretical calculations suggest that the ester linkers, which are in close proximity to one another due to the π-stacking interactions, induce attractive electrostatic forces and augment self-association. The self-assembly of TPT dyes into well-defined 1D clusters with high aspect ratios was observed, and their morphologies and crystallinity were investigated using SEM and X-ray diffraction analyses. TPTs with ester linkers exhibit a columnar liquid crystalline mesophase in the condensed phase.

EXTRAÇÃO DE CORANTES TÊXTEIS UTILIZANDO SISTEMAS AQUOSOS BIFÁSICOS COMPOSTOS POR 2-PROPANOL E SAIS DE SULFATO

EXTRACTION OF TEXTILE DYES USING TWO-PHASE AQUEOUS SYSTEMS COMPOSED OF 2-PROPANOL AND SULFATE SALTS. Aqueous Two Phase Systems (ATPS) composed of 2-propanol + inorganic salt (Na2SO4/MgSO4) + water, at different temperatures T = (20, 30, 40 and 50) °C were applied in the partition studies of the textile dyes Yellow, Blue Royal and Red Dianix CC. The thermodynamic parameters of the free transfer energy (ΔGtr), enthalpy (ΔHtr) and entropy (ΔStr), respectively, were determined for dye partitioning, using the Van’t Hoff equation. The values of the partition coefficients of textile dyes Yellow, Royal Blue and Red Dianix CC, ranged from 14.21 to 1610.00, 15.15 to 994.00 and 17.32 to 311.75, respectively; and with dye extraction yields ranging from 93.17 to 99.93; 94.53 to 99.89 and 97.19 to 99.64, respectively. Based on the ΔGtr results, it was verified that the dyes partition process was thermodynamically spontaneous, in all evaluated systems. For the 2-propanol + sodium sulfate ATPS, there was a prevalence of the enthalpic contribution over the entropic contribution (ΔHtr > ΔStr), while for the 2-propanol + magnesium sulfate systems, a greater predominance of the entropic contribution was observed (ΔHtr < ΔStr). The recovery yields (Y) values obtained were above 90% for all systems.

Investigating the entropic nature of membrane-mediated interactions driving the aggregation of peripheral proteins.

Peripheral membrane-associated proteins are known to accumulate on the surface of biomembranes as a result of membrane-mediated interactions. For a pair of rotationally-symmetric curvature-inducing proteins, membrane mechanics at the low-temperature limit predicts pure repulsion. On the other hand, temperature-dependent entropic forces arise between pairs of stiff-binding proteins suppressing membrane fluctuations. These Casimir-like interactions have thus been suggested as candidates for attractive forces leading to aggregation. With dense assemblies of peripheral proteins on the membrane, both these abstractions encounter short-range and multi-body complications. Here, we make use of a particle-based membrane model augmented with flexible peripheral proteins to quantify purely membrane-mediated interactions and investigate their underlying nature. We introduce a continuous reaction coordinate corresponding to the progression of protein aggregation. We obtain free energy and entropy landscapes for different surface concentrations along this reaction coordinate. In parallel, we investigate time-dependent estimates of membrane entropy corresponding to membrane undulations and coarse-grained director field and how they change dynamically with protein aggregation. Congruent outcomes of the two approaches point to the conclusion that for low surface concentrations, interactions with an entropic nature may drive the aggregation. But at high concentrations, enthalpic contributions due to concerted membrane deformation by protein clusters are dominant.

Thermodynamics of Pillararene•Guest Complexation: Blinded Dataset for the SAMPL9 Challenge.

We report an investigation of the complexation between a water soluble pillararene host (WP6) and a panel of hydrophobic cationic guests (G1 - G20) by a combination of 1H NMR spectroscopy and isothermal titration calorimetry in phosphate buffered saline. We find that WP6 forms 1:1 complexes with Ka values in the 104 - 109 M-1 range driven by favorable enthalpic contributions. This thermodynamic dataset serves as blinded data for the SAMPL9 challenge.

Characterizing Anion Adsorption to Aqueous Interfaces: Toluene–Water versus Air–Water

We continue our investigation of the behavior of simple ions at aqueous interfaces, employing the combination of two surface-sensitive nonlinear spectroscopy tools, broadband deep UV electronic sum-frequency generation and UV second harmonic generation, to characterize the adsorption of thiocyanate to the interface of water with toluene─a prototypical hydrophobe. We find that both the interfacial spectrum and the Gibbs free energy of adsorption closely match results previously reported for the air-water interface. We observe no relative spectral shift in the higher-energy CTTS transition of thiocyanate, implying similar solvation environments for the two interfaces. Similarly, the Gibbs free energies of adsorption agree within error; however, we expect the respective enthalpic and entropic contributions to differ between the two interfaces, similar to our earlier findings for the air-water versus graphene-water interfaces. Further experiments and theoretical modeling are necessary to quantify the mechanistic differences.

Lysine-Dependent Entropy Effects in the B. subtilis Lysine Riboswitch: Insights from Single-Molecule Thermodynamic Studies.

Riboswitches play an important role in RNA-based sensing/gene regulation control for many bacteria. In particular, the accessibility of multiple conformational states at physiological temperatures allows riboswitches to selectively bind a cognate ligand in the aptamer domain, which triggers secondary structural changes in the expression platform, and thereby "switching" between on or off transcriptional or translational states for the downstream RNA. The present work exploits temperature-controlled, single-molecule total internal reflection fluorescence (TIRF) microscopy to study the thermodynamic landscape of such ligand binding/folding processes, specifically for the Bacillus subtilis lysine riboswitch. The results confirm that the riboswitch folds via an induced-fit (IF) mechanism, in which cognate lysine ligand first binds to the riboswitch before structural rearrangement takes place. The transition state to folding is found to be enthalpically favored (ΔHfold‡ < 0), yet with a free-energy barrier that is predominantly entropic (-TΔSfold‡ > 0), which results in folding (unfolding) rate constants strongly dependent (independent) of lysine concentration. Analysis of the single-molecule kinetic "trajectories" reveals this rate constant dependence of kfold on lysine to be predominantly entropic in nature, with the additional lysine conferring preferential advantage to the folding process by the presence of ligands correctly oriented with respect to the riboswitch platform. By way of contrast, van't Hoff analysis reveals enthalpic contributions to the overall folding thermodynamics (ΔH0) to be surprisingly constant and robustly independent of lysine concentration. The results demonstrate the crucial role of hydrogen bonding between the ligand and riboswitch platform but with only a relatively modest fraction (45%) of the overall enthalpy change needed to access the transition state and initiate transcriptional switching.

Concentration dependence of the bulk and surface properties of liquid AlZn alloys at 1000 K

The properties of mixing of the segregating liquid AlZn alloys at 1000 K have been investigated using the joint effect of size ratio and enthalpic contribution as well as entropic effect. The theoretical values of the bulk properties i.e. thermodynamic properties such as excess free energy of mixing, free energy of mixing, entropy of mixing, heat of mixing as well as the structural properties like concentration-concentration partial structure factor (or concentration fluctuations in the long wavelength limit) and short-range order parameter have been compared with the corresponding experimental data available in the literature. Again Butler model has been applied to study the surface properties such as the surface concentration, surface tension and surface concentration-concentration partial structure factor (or surface concentration fluctuations) of liquid AlZn alloys at 1000 K. The theoretical data of the surface tension have been compared with the experimental data available in the literature which show a qualitative agreement.

Effects of Molecular Crowders on Single-Molecule Nucleic Acid Folding: Temperature-Dependent Studies Reveal True Crowding vs Enthalpic Interactions.

Biomolecular folding in cells can be strongly influenced by spatial overlap/excluded volume interactions (i.e., "crowding") with intracellular solutes. As a result, traditional in vitro experiments with dilute buffers may not accurately recapitulate biomolecule folding behavior in vivo. In order to account for such ubiquitous excluded volume effects, biologically inert polyethylene glycol (PEG) and polysaccharides (dextran and Ficoll) are often used as in vitro crowding agents to mimic in vivo crowding conditions, with a common observation that high concentrations of these polymers stabilize the more compact biomolecule conformation. However, such an analysis can be distorted by differences in polymer interactions with the folded vs unfolded conformers, requiring temperature-dependent analysis of the thermodynamics to reliably assess competing enthalpic vs entropic contributions and thus the explicit role of excluded volume. In this work, temperature-controlled single-molecule fluorescence resonance energy transfer (smFRET) is used to characterize the thermodynamic interaction between nucleic acids and common polymer crowders PEG, dextran, and Ficoll. The results reveal that PEG promotes secondary and tertiary nucleic acid folding by simultaneously increasing the folding rate while decreasing the unfolding rate, with temperature-dependent studies confirming that the source of PEG stabilization is predominantly entropic and consistent with a true excluded volume crowding mechanism. By way of contrast, neither dextran nor Ficoll induces any significant concentration-dependent change in nucleic acid folding stability at room temperature, but instead, stabilization effects gradually appear with a temperature increase. Such a thermal response indicates that both folding enthalpies and entropies are impacted by dextran and Ficoll. A detailed thermodynamic analysis of the kinetics suggests that, instead of true entropic molecular crowding, dextran and Ficoll associate preferentially with the unfolded vs folded nucleic acid conformer as a result of larger solvent accessible surface area, thereby skewing the free energy landscapes through both significant entropic/enthalpic contributions that compete and fortuitously cancel near room temperature.