Water-macromolecule Interactions Research Articles

Ion-Specific Water–Macromolecule Interactions at the Air/Aqueous Interface: An Insight into Hofmeister Effect

The specificity of ions in inducing conformational changes in macromolecules is introduced as the Hofmeister series; however, the detailed underlying mechanism is not comprehensible yet. We utilized surface-specific sum frequency generation (SFG) vibrational spectroscopy to explore the Hofmeister effect at the air/polyvinylpyrrolidone (PVP)/water interface. The spectral signature observed from the ssp polarization scheme reveals ion-specific ordering of water molecules following the Hofmeister series attributed to the ion-macromolecule interactions. Along with this, the presence of ions does not reflect any significant influence on the structure of the PVP macromolecule. However, the ppp-SFG spectra in the CH-stretch region reveal the impact of ions on the orientation angle of vinyl chain CH2-groups, which follows the Hofmeister series: SO42- > Cl- > NO3- > Br- > ClO4- > SCN-. The minimal orientation angle of CH2-groups indicates significant reordering in PVP vinyl chains in the presence of chaotropic anions ClO4-, and SCN-. The observation is attributed to the ion-specific water-macromolecule interactions at the air/aqueous interface. It is compelling to observe the signature of spectral blue shifts in the OH-stretch region in the ppp configuration in the presence of chaotropic anions. The origin of spectral blue shifts has been ascribed to the existence of weaker interactions between the interfacial water molecules and the backbone CH- and CH2-moieties of the PVP macromolecules. The ion-specific modulation in water-macromolecule interactions is endorsed by the relative propensity of anion's adsorption toward the air/aqueous interface. The experimental findings highlight the existence and cooperative participation of ion-specific water-macromolecule interactions in the mechanism of the Hofmeister effect, along with the illustrious ion-water and ion-macromolecule interactions.

Microstructure and water distribution of commercial pasta studied by microscopy and 3D magnetic resonance imaging

Manufacturing pasta is a rather well known process, but it is still challenging to tailor pasta products with new raw materials. In this study, we evaluated the effects of raw materials on the microstructure and water distribution in cooked pasta using 1H magnetic resonance imaging (MRI) as well as bright field and polarized light microscopy. The MRI parameters initial intensity (I0) and transverse dephasing time (T2⁎) serve as indicators of the local water concentration and water–macromolecule interactions through chemical exchange, respectively. These parameters were mapped throughout the whole pasta volume with a spatial resolution of 78μm in all three dimensions. MRI was combined with light microscopy to link I0 and T2⁎ to microstructure components such as fiber particles and the extent of starch gelatinization. Four commercial spaghetti samples were analyzed which were made of durum wheat flour, both plain and enriched with wheat fiber, as well as with wholegrain and soft wheat flour. Although all pasta samples showed similar macroscopic water absorption as measured by weight increase, the sample structures differed at the microscopic scale. Compared to durum wheat spaghetti, the presence of fiber particles decreased T2⁎, while spaghetti enriched with soft wheat flour increased T2⁎. In addition, light microscopy showed that large fiber particles partly acted as barriers against water migration and protected starch granules from swelling. Smaller wheat fiber particles did not affect local starch swelling. Thus, the combination of light microscopy and MRI is a powerful tool to study the microstructure and water distribution in pasta.

Water-Protein Interactions: The Secret of Protein Dynamics

Water-protein interactions help to maintain flexible conformation conditions which are required for multifunctional protein recognition processes. The intimate relationship between the protein surface and hydration water can be analyzed by studying experimental water properties measured in protein systems in solution. In particular, proteins in solution modify the structure and the dynamics of the bulk water at the solute-solvent interface. The ordering effects of proteins on hydration water are extended for several angstroms. In this paper we propose a method for analyzing the dynamical properties of the water molecules present in the hydration shells of proteins. The approach is based on the analysis of the effects of protein-solvent interactions on water protons NMR relaxation parameters. NMR relaxation parameters, especially the nonselective (R 1 NS ) and selective (R 1 SE ) spin-lattice relaxation rates of water protons, are useful for investigating the solvent dynamics at the macromolecule-solvent interfaces as well as the perturbation effects caused by the water-macromolecule interactions on the solvent dynamical properties. In this paper we demonstrate that Nuclear Magnetic Resonance Spectroscopy can be used to determine the dynamical contributions of proteins to the water molecules belonging to their hydration shells.

Water–water and water–macromolecule interactions in food dehydration and the effects of the pore structures of food on the energetics of the interactions

A molecular dynamics (MD) modeling and simulations approach has been rationally built and developed to study porous food systems constructed with amylose and dextran chains. The findings from our MD studies indicate that the presence of food macromolecules decreases the energetics of the water–water interactions for the nearby water molecules in the pore space, but provides additional water–macromolecule interactions that can significantly outweigh the partial loss of water–water interactions to make the adjacent water molecules strongly bound to the food macromolecules so that the water activity and water removal rate are decreased as dehydration proceeds and, thus, the dehydration energy requirement would be increased. The effects of pore structures are greater in systems with higher densities of food macromolecules, smaller in size pores, and stronger water–macromolecule interactions. Dehydration of food materials can thus be reasonably expected to start from the largest pores and from the middle of the pores, and to have non-uniform water removal rates and non-planar water–vapor interfaces inside individual pores as well as across sections of the food materials. The food porous structures are found to have good pore connectivity for water molecules. As dehydration proceeds, water content and the support from water–water and water–macromolecule interactions both decrease, causing the food porous structures to adopt more compact conformations and their main body to decrease in size. Dehydration in general also reduces pore sizes and the number of pore openings, increases the water–macromolecule interactions, and leads to the reduction of the overall thermal conductivity of the system, so that more energy (heat), longer times, and/or greater temperature gradients are needed in order to further dehydrate the porous materials. Our thermodynamic analysis also shows that the average minimum entropy requirement for food dehydration is greater when the water–macromolecule interactions are stronger and the food macromolecular density is higher. The importance of the physicochemical affinity of food molecules for water and of the compatibility of the resultant porous structures with water configurational structures in determining food properties and food processing through the water–macromolecule interactions, is clearly and fundamentally verified by the results and discussion presented in this work.

Z-spectroscopy with Alternating-Phase Irradiation

Magnetization transfer (MT) MRI and Z-spectroscopy are tools to study both water–macromolecule interactions and pH-sensitive exchange dynamics between water and the protons of mobile chemical groups within these macromolecules. Both rely on saturation of frequencies offset from water and observation of the on-resonance water signal. In this work, an RF saturation method called Z-spectroscopy with Alternating-Phase Irradiation (ZAPI) is introduced. Based on the T 2-selectivity of the irradiation pulse, ZAPI can be used to separate the different contributions to a Z-spectrum, as well as to study the T 2 distribution of the macromolecules contributing to the MT signal. ZAPI can be run at resonance for water and with low power, thus minimizing problems with specific absorption rate (SAR) limits in clinical applications. In this paper, physical and practical aspects of ZAPI are discussed and the sequence is applied in vitro to sample systems and in vivo to rat head to demonstrate the method.

Investigating the effect of blood susceptibility on phase contrast in the human brain

Recent work has shown a dramatic contrast between GM and WM in gradient echo phase images at high field (7 T). Although this contrast is key to the exploitation of phase in imaging normal and pathological tissue, its origin remains contentious. Several sources for this contrast have been considered including iron content, myelin, deoxy-hemoglobin, or water–macromolecule interactions. Here we quantify the contribution of intravascular dHb to the GM/WM contrast in the human brain at 7 T by modulating the susceptibility of the blood using a paramagnetic contrast agent. By carrying out high resolution, dynamic, gradient echo imaging before, during and after the injection of the contrast agent, we were able to follow the change in GM/WM phase contrast and to monitor simultaneously the susceptibility of the blood. Using these data in conjunction with the known susceptibility of venous blood we estimate the upper bound for the relative contribution of dHb in the vasculature to the measured GM/WM phase contrast to be 0.48 Hz for GM close to the pial surface, and 0.27 Hz for deeper GM. These values are up to 20% of the GM/WM phase difference observed in the human brain at 7 T. Furthermore, we found that the fractional blood volume differences required to account for the observed GM/WM phase contrast are 1.3% and 0.7% for GM close to the pial surface and for deeper GM, respectively.

Magnetic resonance imaging studies of water interactions in meat

The use of MRI shows spatial resolution of water content, and NMR parameters including relaxation times ( T 1 and T 2), and diffusion coefficients ( D) define the state of water interactions with other molecules. These parameters are potentially sensitive to local variations of water mobility and result from modification of water–macromolecule interactions and changes in tissue structure. MRI gives a unique opportunity to better-understand the dynamic phenomena that occur during processing and storage of food. By measuring apparent diffusion coefficient, both axially and radially in meat, it is possible to probe the influence of intracellular diffusional barriers or post-mortem structural changes. The effects of different freezing methods on trout muscle were investigated using MRI. The variations of the relaxation time, T 2, and the radial diffusion coefficient, characterize the structural changes of tissue produced by the freezing process.

4.0 T Water Proton T1 Relaxation Times in Normal Human Brain and During Acute Ethanol Intoxication

It has been reported that acute ethanol intoxication decreases the brain water proton magnetic resonance T1 values, an effect that has been interpreted to indicate brain dehydration during this condition. Because water macromolecular interactions largely determine tissue water T1, another possible explanation for reduced brain water proton T1 values is that the interaction between water and brain macromolecules is altered by ethanol. A 4.0 T magnetic resonance imaging (MRI) instrument was used to measure brain water proton T1 relaxation times before, during, and after ethanol intoxication (dose, 0.75 mg/kg) in healthy controls. The T1 relaxation times as assessed with MRI were highly reproducible. The mean, paired ethanol-induced differences in T1 were -0.004 +/- 0.007 sec (mean +/- standard deviation) for white matter and 0.010 +/- 0.015 sec for internal gray matter structures, neither of which was significant. This reasonably sensitive measurement does not support the view that tissue water content or water macromolecule interactions are significantly altered in the brain during acute alcohol intoxication in otherwise healthy subjects.

4.0 T Water Proton T1 Relaxation Times in Normal Human Brain and During Acute Ethanol Intoxication

Background: It has been reported that acute ethanol intoxication decreases the brain water proton magnetic resonance T1 values, an effect that has been interpreted to indicate brain dehydration during this condition. Because water macromolecular interactions largely determine tissue water T1, another possible explanation for reduced brain water proton T1 values is that the interaction between water and brain macromolecules is altered by ethanol. Methods: A 4.0 T magnetic resonance imaging (MRI) instrument was used to measure brain water proton T1 relaxation times before, during, and after ethanol intoxication (dose, 0.75 mg/kg) in healthy controls. Results: The T1 relaxation times as assessed with MRI were highly reproducible. The mean, paired ethanol-induced differences in T1 were −0.004 ± 0.007 sec (mean ± standard deviation) for white matter and 0.010 ± 0.015 sec for internal gray matter structures, neither of which was significant. Conclusions: This reasonably sensitive measurement does not support the view that tissue water content or water macromolecule interactions are significantly altered in the brain during acute alcohol intoxication in otherwise healthy subjects.

Water-macromolecule interactions by NMR: a quadrature-free constant-time approach and its application to CI2.

A pulse sequence is proposed to select water magnetization with enhanced specificity through a synergetic combination of several filtering principles. This approach relies on a constant-time evolution period implemented without quadrature detection, which results in a square root 2 increase in signal-to-noise ratios as compared to traditional non-selective methods for water filtration. In addition, the quadrature-free constant-time block facilitates the implementation of the water flip-back strategy, which leads to further gains in sensitivity. The proposed experiment was applied to unlabeled HEW lysozyme and to 15N-labeled chymotrypsin inhibitor 2 which was partially or non 13C-enriched. Water molecules belonging to a spine of hydration between two pseudo beta-sheet strands were identified, solving previously reported discrepancies between the X-ray and refined NMR structure of CI2. The proposed experiment in particularly suitable for hydration studies of mixtures of labeled and unlabeled components, such as ligand-macromolecule complexes.

Band-selective editing of exchange-relay in protein-water NOE experiments.

A pulse sequence is proposed which uses a train of band-selective pulses for the editing of slow chemical exchange-relay effects in experiments designed to study water-macromolecule interactions. Compared to previous methods, this experiment does not require knowledge of the exact chemical shift of the relaying labile protons and needs only the recording of a single experiment to edit the relay through different exchanging groups resonating at different frequencies. The pulse sequence has been implemented using Gaussian cascades and was applied to the study of the hydration of HEW lysozyme.

The Role of Specific Side Groups and pH in Magnetization Transfer in Polymers

The nature of water–macromolecule interactions in aqueous model polymers has been investigated using quantitative measurements of magnetization transfer. Cross-linked polymer gels composed of 94% water, 3%N,N′-methylene-bis-acrylamide, and 3% functional monomer (acrylamide, methacrylamide, acrylic acid, methacrylic acid, 2-hydroxyethyl-acrylate, or 2-hydroxyethyl-methacrylate) were studied. Water–macromolecule interactions were modified by varying the pH and specific functional group on the monomer. The magnitudes of the interactions were quantified by measuring the rate of proton nuclear spin magnetization exchange between the polymer matrix and the water. This rate was highly sensitive to the presence of carboxyl side groups on the macromolecule. However, the dependence of the rate on pH was not consistent with simple acid/base-catalyzed chemical exchange, and instead, the data suggest that multiequilibria proton exchange, a wide distribution in surface group pKvalues, and/or a macromolecular structural dependence on pH may play a significant role in magnetization transfer in polymer systems. These model polymer gels afford useful insights into the relevance of chemical composition and chemical dynamics on relaxation in tissues.

The Effects of Cross-Link Density and Chemical Exchange on Magnetization Transfer in Polyacrylamide Gels

The effects of polymer structure and water–macromolecule interactions on proton relaxation in an aqueous model polymer have been investigated using quantitative measurements of magnetization transfer. Polyacrylamide gels composed of 95% water, 5% comonomers acrylamide andN,N′-methylene-bis-acrylamide were studied. The structure and rigidity were varied by changing the cross-linking density of the polymer. The polymer showed a biphasic change in transverse relaxation with increasing cross-linking density which was accompanied by a sudden increase in magnetization transfer above 40% cross linking. This change may be attributed to the formation of rigid domains in the polymer which exhibit solid-like behavior with a shortT2(11 μs) and a Gaussian lineshape. Water–macromolecule interactions were controlled by varying the pH of the gel. At high pH (>8), there was an increase in magnetization transfer and transverse relaxivity consistent with a chemical-exchange-mediated interaction between water protons and the polymer. By analyzing the system as two proton reservoirs coupled by magnetization exchange, the proton populations, intrinsic relaxation rates, and exchange rates were estimated, for different degrees of cross linking and pH. This model affords useful insights into the relevance of both supramolecular structure and chemical exchange on relaxation in tissues.

A near-infrared analysis of water-macromolecule interactions: Hydration and the spectra of aqueous solutions of intact proteins

A procedure utilizing a variable path length absorption cell has made possible the recording of what appear to be the first well-resolved, compensated near-infrared spectra of intact proteins in aqueous solution. Individual spectra, corresponding to (1) absorbance by the protein plus bound water, and (2) the solvent volume excluded by the hydrated protein, were obtained using the same experimental sample. Calculations of bound water and excluded volume from these spectra were compared to other results in the literature. The validity of this spectral method was supported by comparisons with the spectra of proteins in films, where there is no excluded volume effect and where the amount of water present has been determined independently by gravimetric measurements. The results indicate that the bound water detected by the near-infrared spectra has an absence or deficiency of molecules with quasi-free OH groups (relative to bulk water), and that in conjunction with results of other methods, these water molecules may represent those that are most firmly or more completely bonded to the protein surface.