Water and protein dynamics in protein-water mixtures over wide range of composition

© 2012 IEEE. Personal use of this material is permitted. Permission from IEEE must be obtained for all other uses, in any current or future media, including reprinting/republishing this material for advertising or promotional purposes, creating new collective works, for resale or redistribution to servers or lists, or reuse of any copyrighted component of this work in other works.[EN] Water and protein dynamics in two globular protein-water systems, water-lysozyme and water-BSA (bovine serum albumine), were studied by differential scanning calorimetry (DSC), dielectric relaxation spectroscopy (DRS) and thermally stimulated depolarization currents (TSDC) techniques. Water equilibrium sorption isotherms (ESI) measurements were also recorded at room temperature. The samples covered a wide range of composition, from practically dry solid pellets (2wt% of water) to dilute solutions (82wt% of water). Crystallization and melting events of water were studied by DSC and the amount of uncrystallized water was calculated. The evolution of dynamics with hydration level was followed for various dielectric relaxation processes, the emphasis being given to relaxation processes of polar groups on the surface of the proteins and of uncrystallized water molecules. A relationship between the formation of a conductive percolating water cluster and the saturation of the water process was found.This research has been co-financed by the European Union (European Social Fund - ESF) and Greek national funds through the Operational Program "Education and Lifelong Learning" of the National Strategic Reference Framework (NSRF) - Research Funding Program: Heracleitus II. Investing in knowledge society through the European Social Fund.Kyritsis, A.; Panagopoulou, A.; Pissis, P.; Sabater I Serra, R.; Gómez Ribelles, JL.; Shinyashiki, N. (2012). Water and protein dynamics in protein-water mixtures over wide range of composition. IEEE Transactions on Dielectrics and Electrical Insulation. 19(4):1239-1246. https://doi.org/10.1109/TDEI.2012.6259997S1239124619

Recognition of a new permittivity function for glycerol by the use of the eigen-coordinates method

Measurements of real and imaginary parts of the relative complex permittivity of glycerol were carried out in the frequency range 1 mHz-1 MHz at different temperatures between 188 and 263 K. The permittivity data have been analyzed thoroughly by a new data curve-fitting approach that involves the so-called eigen-coordinates method in conjunction with a separation procedure and the inverse permittivity formulas. A new single permittivity function, based on the so-called recap element picture for a self-similar (fractal) structure, has been recognized to describe well such data over the entire frequency range studied. The recognized dielectric function enabled us to infer an electrical equivalent-circuit network for the glycerol sample studied that involves a series combination of two recap elements, reflecting the existence of two different dielectric relaxation processes in glycerol. The temperature dependence of the relaxation times τ1(T) and τ2(T) entering into the identified permittivity function was found to obey nearly an Arrhenius behaviour with activation energies E1 ≈ 114 kJ/mol and E2 ≈ 94 kJ/mol. The recognized permittivity function can be justified by presuming that the processes represented by the recap elements characterized by the parameters (ν1, τ1, E1) and (ν2, τ2, E2) are linked to 'donor-like' and 'acceptor-like' charges formed from the infinite hydroxyl hydrogen bonds. © 2002 Elsevier Science B.V. All rights reserved

HNO Binding in a Heme Protein: Structures, Spectroscopic Properties, and Stabilities

HNO can interact with numerous heme proteins, but atomic level structures are largely unknown. In this work, various structural models for the first stable HNO heme protein complex, MbHNO (Mb, myoglobin), were examined by quantum chemical calculations. This investigation led to the discovery of two novel structural models that can excellently reproduce numerous experimental spectroscopic properties. They are also the first atomic level structures that can account for the experimentally observed high stabilities. These two models involve two distal His conformations as reported previously for MbCNR and MbNO. However, a unique dual hydrogen bonding feature of the HNO binding was not reported before in heme protein complexes with other small molecules such as CO, NO, and O2. These results shall facilitate investigations of HNO bindings in other heme proteins

Evidence of coexistence of change of caged dynamics at Tg and the dynamic transition at Td in solvated proteins

Mossbauer spectroscopy and neutron scattering measurements on proteins embedded in solvents including water and aqueous mixtures have emphasized the observation of the distinctive temperature dependence of the atomic mean square displacements, , commonly referred to as the dynamic transition at some temperature Td. At low temperatures, increases slowly, but it assume stronger temperature dependence after crossing Td, which depends on the time/frequency resolution of the spectrometer. Various authors have made connection of the dynamics of solvated proteins including the dynamic transition to that of glass-forming substances. Notwithstanding, no connection is made to the similar change of temperature dependence of obtained by quasielastic neutron scattering when crossing the glass transition temperature Tg, generally observed in inorganic, organic and polymeric glass-formers. Evidences are presented to show that such change of the temperature dependence of from neutron scattering at Tg is present in hydrated or solvated proteins, as well as in the solvents used unsurprisingly since the latter is just another organic glass-formers. The obtained by neutron scattering at not so low temperatures has contributions from the dissipation of molecules while caged by the anharmonic intermolecular potential at times before dissolution of cages by the onset of the Johari-Goldstein beta-relaxation. The universal change of at Tg of glass-formers had been rationalized by sensitivity to change in volume and entropy of the beta-relaxation, which is passed onto the dissipation of the caged molecules and its contribution to . The same rationalization applies to hydrated and solvated proteins for the observed change of at Tg.Comment: 28 pages, 10 figures, 1 Tabl

Through vial impedance spectroscopy (TVIS): A novel approach to process understanding for freeze-drying cycle development

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link.Through vial impedance spectroscopy (TVIS) provides a new process analytical technology for monitoring a development scale lyophilization process, which exploits the changes in the bulk electrical properties that occur on freezing and subsequent drying of a drug solution. Unlike the majority of uses of impedance spectroscopy, for freeze-drying process development, the electrodes do not contact the product but are attached to the outside of the glass vial which is used to contain the product to provide a non-sample-invasive monitoring technology. Impedance spectra (in frequency range 10 Hz to 1 MHz) are generated throughout the drying cycle by a specially designed impedance spectrometer based on a 1 GΩ trans-impedance amplifier and then displayed in terms of complex capacitance. Typical capacitance spectra have one or two peaks in the imaginary capacitance (i.e., the dielectric loss) and the same number of steps in the real part capacitance (i.e., the dielectric permittivity). This chapter explores the underlying mechanisms that are responsible for these dielectric processes, i.e., the Maxwell-Wagner (space charge) polarization of the glass wall of the vial through the contents of the vial when in the liquid state, and the dielectric relaxation of ice when in the frozen state. In future work, it will be demonstrated how to measure product temperature and drying rates within single vials and multiple (clusters) of vials, from which other critical process parameters, such as heat transfer coefficient and dry layer resistance, may be determined

Universal secondary relaxation of water in aqueous mixtures, in nano-confinement, and in hydrated proteins

From a large volume of experimental data of relaxation of water in various aqueous mixtures, in different forms of nano-confinement, and in two hydrated proteins, we show the presence of a secondary relaxation in all these systems that have similar characteristics. This ubiquitous secondary relaxation originates from water in the systems and is the analogue of the universal Johari-Goldstein secondary relaxation of glass-forming substances in general. Like all Johari-Goldstein secondary relaxation, this one from water bears an intimate and important relation to the primary structural relaxation, and hence it plays a fundamental role in glass transition or function of these water containing systems. © 2008 American Institute of Physics

The protein "glass" transition and the role of the solvent

Hydrated proteins undergo a change in their dynamical properties in the neighborhood of a temperature. The change of dynamics has been likened to glass transition of glass-forming substances because similar properties were found. However, a complete understanding of the conformation fluctuations of hydrated proteins and their relation to the dynamics of the solvent is still not available, possibly due to the protein molecules being more complex than ordinary glass-formers. For this reason, we turn our attention to the experimental findings of the dynamics of mixtures of water with simpler glass-formers (small molecules and polymers). Two major relaxation processes have been observed in these aqueous mixtures. One is the structural alpha-relaxation of the hydrophilic glass-former hydrogen bonded to the water, which is responsible for glass transition. The other one is the local secondary beta-relaxation of water in the mixture. Remarkably, these two relaxation processes in aqueous mixtures have analogues in hydrated proteins with the same properties. The conformation fluctuations of the protein and the relaxation of the solvent in hydrated proteins behave like the alpha-relaxation of the hydrophilic glass-former hydrogen bonded to the water and the beta-relaxation of water in other aqueous mixtures, respectively. At low temperatures, the Arrhenius activation energy of the relaxation time of the solvent in a hydrated protein is almost the same as that of the beta-relaxation of water in the glassy states of aqueous mixtures. The Arrhenius T-dependence of the solvent relaxation times no longer holds at temperatures that exceed the "glass" transition temperature of the hydrated protein, defined as the temperature at which the conformation relaxation time is very long. This behavior of the solvent in hydrated proteins is similar to that found in the beta-relaxation of water in aqueous mixtures when crossing the glass transition temperature of the mixture (Capaccioli, S.; Ngai, K. L.; Shinyashiki, N. J. Phys. Chem. B 2007, 111, 8197). Furthermore, the same dynamics were found in mixtures of two van der Waals glass-formers, which are even simpler systems than aqueous mixtures because of the absence of hydrogen bonding. The experimental data of these ideal mixtures of van der Waals glass-formers have been given a satisfactory theoretical explanation. Since the properties of hydrated proteins, aqueous mixtures, and the mixtures of van der Waals liquids are similar, we transfer the theoretical understanding gained in the study of the last system sequentially to the two other increasingly more complex systems