Preferential solvation of lysozyme in water/ethanol mixtures.

We provide a quantitative description of the solvation properties of lysozyme in water/ethanol mixtures, which has been obtained by a simultaneous analysis of small-angle neutron scattering and differential scanning calorimetry experiments. All data sets were analyzed by an original method, which integrates the exchange equilibrium model between water and ethanol molecules at the protein surface and activity coefficients data of water/ethanol binary mixtures. As a result, the preferential binding of ethanol molecules at the protein surface was obtained for both native and thermal unfolded protein states. Excess solvation numbers reveal a critical point at ethanol molar fraction ≈0.06, corresponding to the triggering of the hydrophobic clustering of alcohol molecules detected in water/ethanol binary mixtures.

[1]  R Diamond,et al.  Real-space refinement of the structure of hen egg-white lysozyme. , 1977, Journal of molecular biology.

[2]  S. Cinelli,et al.  Conformational changes in the unfolding process of lysozyme in water and ethanol/water solutions , 2011 .

[3]  J. Schellman Protein stability in mixed solvents: a balance of contact interaction and excluded volume. , 2003, Biophysical journal.

[4]  G. Onori,et al.  Self‐association of monohydric alcohols in water: Compressibility and infrared absorption measurements , 1994 .

[5]  Yun Liu,et al.  Cluster formation in two-Yukawa fluids. , 2005, The Journal of chemical physics.

[6]  Xiaoning Yang,et al.  Molecular dynamics simulation of ethanol/water mixtures for structure and diffusion properties , 2005 .

[7]  Milo M. Lin,et al.  Solvation in protein (un)folding of melittin tetramer–monomer transition , 2009, Proceedings of the National Academy of Sciences.

[8]  Hui Lu,et al.  Solvent molecules bridge the mechanical unfolding transition state of a protein , 2008, Proceedings of the National Academy of Sciences.

[9]  H. Ågren,et al.  Molecular structure of alcohol-water mixtures. , 2003, Physical review letters.

[10]  Eli Ruckenstein,et al.  A protein molecule in an aqueous mixed solvent: fluctuation theory outlook. , 2005, The Journal of chemical physics.

[11]  F. Spinozzi,et al.  Looking for the best experimental conditions to detail the protein solvation shell in a binary aqueous solvent via small angle scattering , 2009 .

[12]  S. Cinelli,et al.  Effect of 1-alcohols on DNA condensation and protein folding , 2010 .

[13]  S. N. Timasheff,et al.  Preferential interactions of urea with lysozyme and their linkage to protein denaturation. , 2003, Biophysical chemistry.

[14]  Rowan Hargreaves,et al.  Clustering and microimmiscibility in alcohol-water mixtures : Evidence from molecular-dynamics simulations , 2005 .

[15]  M. Malfois,et al.  Proteins in solution : from X-ray scattering intensities to interaction potentials , 1999 .

[16]  J. Schellman Destabilization and stabilization of proteins , 2005, Quarterly Reviews of Biophysics.

[17]  T. S. Banipal,et al.  Thermodynamic study of solvation of some amino acids, diglycine and lysozyme in aqueous and mixed aqueous solutions , 2004 .

[18]  Sow-Hsin Chen Small Angle Neutron Scattering Studies of the Structure and Interaction in Micellar and Microemulsion Systems , 1986 .

[19]  M. Wertheim,et al.  EXACT SOLUTION OF THE PERCUS-YEVICK INTEGRAL EQUATION FOR HARD SPHERES , 1963 .

[20]  J. Crain,et al.  Excess entropy in alcohol-water solutions: a simple clustering explanation. , 2006, The journal of physical chemistry. B.

[21]  G. Onori,et al.  Dynamical and structural properties of water/alcohol mixtures , 1996 .

[22]  J. Schellman,et al.  The thermodynamics of solvent exchange , 1994, Biopolymers.

[23]  Frédéric Cardinaux,et al.  Equilibrium cluster formation in concentrated protein solutions and colloids , 2004, Nature.

[24]  D. Clemens,et al.  New insights into urea action on proteins: a SANS study of the lysozyme case. , 2008, The journal of physical chemistry. B.

[25]  R. S. Hansen,et al.  A New Method for Determination of Activities of Binary Solutions of Volatile Liquids , 1954 .

[26]  M. Lehmann,et al.  Neutron small-angle scattering studies of ribonuclease in mixed aqueous solutions and determination of the preferentially bound water. , 1984, Biochemistry.

[27]  F. Spinozzi,et al.  SANS/SAXS study of the BSA solvation properties in aqueous urea solutions via a global fit approach , 2008, European Biophysics Journal.

[28]  T. M. Raschke,et al.  Water structure and interactions with protein surfaces. , 2006, Current opinion in structural biology.

[29]  Valerie Daggett,et al.  The molecular mechanism of stabilization of proteins by TMAO and its ability to counteract the effects of urea. , 2002, Journal of the American Chemical Society.

[30]  A. Ciferri,et al.  Thermodynamics of unfolding of lysozyme in aqueous alcohol solutions. , 1973, The Journal of biological chemistry.

[31]  G. Onori,et al.  Examination by dynamic light scattering of lysozyme in water/alcohol mixtures , 2001 .

[32]  P. Schurtenberger,et al.  Cluster-driven dynamical arrest in concentrated lysozyme solutions. , 2011, The journal of physical chemistry. B.

[33]  Á. Domínguez,et al.  Density, dynamic viscosity, and derived properties of binary mixtures of methanol or ethanol with water, ethyl acetate, and methyl acetate at T=(293.15, 298.15, and 303.15) K , 2007 .

[34]  J. Grolier,et al.  Excess volumes and excess heat capacities of water + ethanol at 298.15 K , 1981 .

[35]  F. Spinozzi,et al.  Preferential hydration of lysozyme in water/glycerol mixtures: a small-angle neutron scattering study. , 2007, The Journal of chemical physics.

[36]  Paolo Mariani,et al.  Interaction of proteins in solution from small-angle scattering: a perturbative approach. , 2002, Biophysical journal.

[37]  B. Berne,et al.  Dewetting and hydrophobic interaction in physical and biological systems. , 2009, Annual review of physical chemistry.

[38]  J. Teixeira Introduction to Small Angle Neutron Scattering Applied to Colloidal Science , 1992 .

[39]  S. N. Timasheff,et al.  Protein-solvent preferential interactions, protein hydration, and the modulation of biochemical reactions by solvent components , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[40]  D. Svergun,et al.  Absence of equilibrium cluster phase in concentrated lysozyme solutions , 2008, Proceedings of the National Academy of Sciences.

[41]  P. Baglioni,et al.  Lysozyme protein solution with an intermediate range order structure. , 2011, The journal of physical chemistry. B.

[42]  F. Spinozzi,et al.  Combining structure and dynamics: non-denaturing high-pressure effect on lysozyme in solution , 2009, Journal of The Royal Society Interface.

[43]  S. Suresh,et al.  Theory of dielectric constant of aqueous solutions , 2002 .

[44]  P. Baglioni,et al.  The two-Yukawa model and its applications: the cases of charged proteins and copolymer micellar solutions , 2007 .

[45]  F. Spinozzi,et al.  Microcalorimetric study of thermal unfolding of lysozyme in water/glycerol mixtures: an analysis by solvent exchange model. , 2008, The Journal of chemical physics.

[46]  A. Wakisaka,et al.  Phase separation of water-alcohol binary mixtures induced by the microheterogeneity. , 2005, Faraday discussions.

[47]  E. Castner,et al.  A molecular dynamics study of aggregation phenomena in aqueous n-propanol , 2004 .

[48]  P. Wolynes,et al.  The energy landscapes and motions of proteins. , 1991, Science.

[49]  K. Gekko,et al.  Mechanism of protein stabilization by glycerol: preferential hydration in glycerol-water mixtures. , 1981, Biochemistry.

[50]  A. Pastore,et al.  Cold denaturation of yeast frataxin offers the clue to understand the effect of alcohols on protein stability. , 2008, Journal of the American Chemical Society.

[51]  R. Jansen,et al.  Ethanol-perturbed amyloidogenic self-assembly of insulin: looking for origins of amyloid strains. , 2005, Biochemistry.

[52]  A. G. Williamson,et al.  Enthalpies of mixture of ethanol and water at 25.degree.C , 1967 .

[53]  Yun Liu,et al.  The structural properties of a two-Yukawa fluid: Simulation and analytical results. , 2006, The Journal of chemical physics.

[54]  J. Prausnitz,et al.  Effect of alcohols on aqueous lysozyme-lysozyme interactions from static light-scattering measurements. , 2004, Biophysical chemistry.