A practical guide on how osmolytes modulate macromolecular properties.

Osmolytes are a class of compounds ubiquitously used by living organisms to respond to cellular stress or to fine-tune molecular properties in the cell. These compounds are also highly useful in vitro. In this chapter, we give an overview of the possible uses of osmolytes in the laboratory, and how we can investigate and understand their modes of action. Experimental procedures are discussed with a specific emphasis on osmolyte-related aspects and on the theoretical aspects that are important to both introductory and more advanced interpretations of such experiments.

[1]  V. Adrian Parsegian,et al.  DNA‐Inspired Electrostatics , 2000 .

[2]  V. Parsegian,et al.  Osmotic stress for the direct measurement of intermolecular forces. , 1986, Methods in enzymology.

[3]  Paul E. Smith Local chemical potential equalization model for cosolvent effects on biomolecular equilibria , 2004 .

[4]  G. Makhatadze THERMODYNAMICS OF PROTEIN INTERACTIONS WITH UREA AND GUANIDINIUM HYDROCHLORIDE , 1999 .

[5]  S. Weerasinghe,et al.  Cavity formation and preferential interactions in urea solutions: Dependence on urea aggregation , 2003 .

[6]  M. Chaudhury,et al.  Solubility of proteins , 1986 .

[7]  C. Tanford,et al.  Proteins as random coils. IV. Osmotic pressures, second virial coefficients, and unperturbed dimensions in 6 M guanidine hydrochloride. , 1967, Journal of the American Chemical Society.

[8]  A. Green STUDIES IN THE PHYSICAL CHEMISTRY OF THE PROTEINS X. THE SOLUBILITY OF HEMOGLOBIN IN SOLUTIONS OF CHLORIDES AND SULFATES OF VARYING CONCENTRATION , 1932 .

[9]  S. Smith‐Gill,et al.  Involvement of water molecules in the association of monoclonal antibody HyHEL-5 with bobwhite quail lysozyme. , 1997, Biophysical journal.

[10]  S. N. Timasheff,et al.  The control of protein stability and association by weak interactions with water: how do solvents affect these processes? , 1993, Annual review of biophysics and biomolecular structure.

[11]  M. Record,et al.  Salt-nucleic acid interactions. , 1995, Annual review of physical chemistry.

[12]  S. Cavagnero,et al.  Kinetic role of electrostatic interactions in the unfolding of hyperthermophilic and mesophilic rubredoxins. , 1998, Biochemistry.

[13]  A. Aggarwal,et al.  Structure and function of restriction endonucleases. , 1995, Current opinion in structural biology.

[14]  T. Kiefhaber,et al.  Kinetic coupling between protein folding and prolyl isomerization. I. Theoretical models. , 1992, Journal of molecular biology.

[15]  M. Record,et al.  Partitioning of atmospherically relevant ions between bulk water and the water/vapor interface , 2006, Proceedings of the National Academy of Sciences.

[16]  J. Brandts,et al.  Determination of the volumetric properties of proteins and other solutes using pressure perturbation calorimetry. , 2002, Analytical biochemistry.

[17]  J. Schellman Solvent denaturation , 1978 .

[18]  D. W. Bolen,et al.  Osmolyte-driven contraction of a random coil protein. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  K. Jung,et al.  Organic osmolyte distribution and levels in the mammalian urinary bladder in diuresis and antidiuresis. , 1996, The American journal of physiology.

[20]  V. Parsegian,et al.  [3] Macromolecules and water: Probing with osmotic stress , 1995 .

[21]  M. Record,et al.  Vapor pressure osmometry studies of osmolyte-protein interactions: implications for the action of osmoprotectants in vivo and for the interpretation of "osmotic stress" experiments in vitro. , 2000, Biochemistry.

[22]  W. Kauzmann,et al.  Pressure denaturation of metmyoglobin. , 1973, Biochemistry.

[23]  Paul E Smith,et al.  Chemical potential derivatives and preferential interaction parameters in biological systems from Kirkwood-Buff theory. , 2006, Biophysical journal.

[24]  K. Dill,et al.  Short-range interactions: from simple ions to polyelectrolyte solutions , 2004 .

[25]  D. G. Archer Thermodynamic Properties of the NaCl+H2O System l. Thermodynamic Properties of NaCl(cr) , 1992 .

[26]  D. Rau,et al.  Exclusion of alcohols from spermidine-DNA assemblies: probing the physical basis of preferential hydration. , 2004, Biochemistry.

[27]  W. Gelbart,et al.  Structure, stability, and thermodynamics of lamellar DNA-lipid complexes. , 1998, Biophysical journal.

[28]  S. N. Timasheff Water as ligand: preferential binding and exclusion of denaturants in protein unfolding. , 1992, Biochemistry.

[29]  T. Lohman,et al.  Thermodynamic extent of counterion release upon binding oligolysines to single-stranded nucleic acids. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[30]  R. A. Goldbeck,et al.  The effect of water on the rate of conformational change in protein allostery. , 2001, Biophysical journal.

[31]  Abu Nayeem Mohammad Salahuddin,et al.  Proteins as random coils. 3. Optical rotatory dispersion in 6 M guanidine hydrochloride. , 1967, Journal of the American Chemical Society.

[32]  Oxana V. Galzitskaya,et al.  Physics of protein folding , 2004 .

[33]  W. Shaw,et al.  The Decomposition of Urea in Aqueous Media , 1955 .

[34]  S. Abdulnur,et al.  EFFECT OF WATER AND OTHER SOLVENTS ON THE STRUCTURE OF BIOPOLYMERS. , 1965, Federation proceedings.

[35]  J. Friedman,et al.  Probing Solvation-Shell Hydrogen Binding in Glassy and Sol−Gel Matrixes through Vibronic Sideband Luminescence Spectroscopy , 2004 .

[36]  M. Record,et al.  Thermodynamics of interactions of urea and guanidinium salts with protein surface: Relationship between solute effects on protein processes and changes in water‐accessible surface area , 2001, Protein science : a publication of the Protein Society.

[37]  K. Hodgson,et al.  Protein denaturation: a small-angle X-ray scattering study of the ensemble of unfolded states of cytochrome c. , 1998, Biochemistry.

[38]  E. Ruckenstein,et al.  A protein molecule in a mixed solvent: the preferential binding parameter via the Kirkwood-Buff theory. , 2006, Biophysical journal.

[39]  D. W. Bolen Effects of naturally occurring osmolytes on protein stability and solubility: issues important in protein crystallization. , 2004, Methods.

[40]  M. Record,et al.  Preferential interactions of glycine betaine and of urea with DNA: implications for DNA hydration and for effects of these solutes on DNA stability. , 2004, Biochemistry.

[41]  Daniel Harries,et al.  Direct Evidence for Counterion Release upon Cationic Lipid−DNA Condensation , 2000 .

[42]  R. Ghirlando,et al.  Probing protein-sugar interactions. , 2000, Biophysical journal.

[43]  H. Hinz,et al.  Response functions of proteins. , 2000, Biophysical chemistry.

[44]  J M Sturtevant,et al.  Thermodynamics of the binding of L-arabinose and of D-galactose to the L-arabinose-binding protein of Escherichia coli. , 1983, The Journal of biological chemistry.

[45]  J. Ferreon,et al.  Protein phase diagrams II: nonideal behavior of biochemical reactions in the presence of osmolytes. , 2007, Biophysical journal.

[46]  G. Vesnaver,et al.  Thermodynamic stability of ribonuclease A in alkylurea solutions and preferential solvation changes accompanying its thermal denaturation: A calorimetric and spectroscopic study , 2008, Protein science : a publication of the Protein Society.

[47]  J. Setschenow Über die Konstitution der Salzlösungen auf Grund ihres Verhaltens zu Kohlensäure , 1889 .

[48]  D. W. Bolen,et al.  Effects of guanidine hydrochloride on the proton inventory of proteins: implications on interpretations of protein stability. , 2000, Biochemistry.

[49]  D. Harries,et al.  Measured depletion of ions at the biomembrane interface. , 2005, Journal of the American Chemical Society.

[50]  C. Tanford,et al.  Proteins as Random Coils. I. Intrinsic Viscosities and Sedimentation Coefficients in Concentrated Guanidine Hydrochloride , 1967 .

[51]  Franz Hofmeister,et al.  Zur Lehre von der Wirkung der Salze , 1891, Archiv für experimentelle Pathologie und Pharmakologie.

[52]  J. Friedman,et al.  Ordered water molecules as key allosteric mediators in a cooperative dimeric hemoglobin. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[53]  B. Lee,et al.  The interpretation of protein structures: estimation of static accessibility. , 1971, Journal of molecular biology.

[54]  Charles Tanford,et al.  The Location of Electrostatic Charges in Kirkwood's Model of Organic Ions , 1957 .

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

[56]  M. Record,et al.  Preferential interactions in aqueous solutions of urea and KCl. , 2003, Biophysical chemistry.

[57]  M M Santoro,et al.  A test of the linear extrapolation of unfolding free energy changes over an extended denaturant concentration range. , 1992, Biochemistry.

[58]  A. C. Ferreon,et al.  Metrics that differentiate the origins of osmolyte effects on protein stability: a test of the surface tension proposal. , 2006, Journal of molecular biology.

[59]  B. Ninham,et al.  The present state of affairs with Hofmeister effects , 2004 .

[60]  M. Stockhausen,et al.  Spectroscopic evidence for the preferential hydration of RNase A in glycerol–water mixtures: Dielectric relaxation studies , 2001 .

[61]  J. Bond,et al.  Thermodynamic characterization of interactions of native bovine serum albumin with highly excluded (glycine betaine) and moderately accumulated (urea) solutes by a novel application of vapor pressure osmometry. , 1996, Biochemistry.

[62]  R. Berisio,et al.  The effect of the osmolyte trimethylamine N‐oxide on the stability of the prion protein at low pH , 2006, Biopolymers.

[63]  M. Burg,et al.  Activity of the TonEBP/OREBP transactivation domain varies directly with extracellular NaCl concentration , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[64]  T. Arakawa,et al.  Theory of protein solubility. , 1985, Methods in enzymology.

[65]  V. Parsegian,et al.  Direct measurement of temperature-dependent solvation forces between DNA double helices. , 1992, Biophysical journal.

[66]  B. Pettitt,et al.  Access the most recent version at doi: 10.1110/ps.062671607 References , 2006 .

[67]  Lu-Yun Lian,et al.  A simple method for improving protein solubility and long-term stability. , 2004, Journal of the American Chemical Society.

[68]  S. Gruner,et al.  Membrane curvature, lipid segregation, and structural transitions for phospholipids under dual-solvent stress. , 1990, Biochemistry.

[69]  Daumantas Matulis,et al.  Thermodynamic stability of carbonic anhydrase: measurements of binding affinity and stoichiometry using ThermoFluor. , 2005, Biochemistry.

[70]  K. Kiyosawa Theoretical and experimental studies on freezing point depression and vapor pressure deficit as methods to measure osmotic pressure of aqueous polyethylene glycol and bovine serum albumin solutions. , 2003, Biophysical chemistry.

[71]  D. Volkin,et al.  Degradative covalent reactions important to protein stability , 1997, Molecular biotechnology.

[72]  Rau,et al.  Measured entropy and enthalpy of hydration as a function of distance between DNA double helices. , 1991, Physical review. A, Atomic, molecular, and optical physics.

[73]  K. Dill,et al.  Charge effects on folded and unfolded proteins. , 1990, Biochemistry.

[74]  J. Schellman Selective binding and solvent denaturation , 1987, Biopolymers.

[75]  D. Goldenberg Computational simulation of the statistical properties of unfolded proteins. , 2003, Journal of molecular biology.

[76]  A. Ben-Naim Theory of preferential solvation of nonelectrolytes , 2008, Cell Biophysics.

[77]  T. Oas,et al.  Linked folding and anion binding of the Bacillus subtilis ribonuclease P protein. , 2001, Biochemistry.

[78]  B. Pettitt,et al.  Uncovering the basis for nonideal behavior of biological molecules. , 2004, Biochemistry.

[79]  J. Friedman,et al.  Molecular Level Probing of Preferential Hydration and Its Modulation by Osmolytes through the Use of Pyranine Complexed to Hemoglobin* , 2006, Journal of Biological Chemistry.

[80]  Ashutosh Tripathy,et al.  Effects of molecular crowding by saccharides on α‐chymotrypsin dimerization , 2002, Protein science : a publication of the Protein Society.

[81]  H. Eisenberg,et al.  THERMODYNAMIC ANALYSIS OF MULTICOMPONENT SOLUTIONS. , 1964, Advances in protein chemistry.

[82]  C. Tanford,et al.  Isopiestic compositions as a measure of preferential interactions of macromolecules in two-component solvents. Application to proteins in concentrated aqueous cesium chloride and guanidine hydrochloride. , 1967, Journal of the American Chemical Society.

[83]  Kevin L. Shaw,et al.  Linear extrapolation method of analyzing solvent denaturation curves , 2000, Proteins.

[84]  H. Hinz,et al.  Dielectric relaxation of aqueous solutions of ribonuclease A in the absence and presence of urea , 1997 .

[85]  G D Rose,et al.  Modeling unfolded states of peptides and proteins. , 1995, Biochemistry.

[86]  Hoang T. Tran,et al.  Reconciling observations of sequence-specific conformational propensities with the generic polymeric behavior of denatured proteins. , 2005, Biochemistry.

[87]  B. Pettitt,et al.  Statistical Thermodynamic Approach to the Chemical Activities in Two-Component Solutions , 2004 .

[88]  A. C. Ferreon,et al.  Thermodynamics of denaturant-induced unfolding of a protein that exhibits variable two-state denaturation. , 2004, Biochemistry.

[89]  M. Burg,et al.  Renal medullary organic osmolytes. , 1991, Physiological reviews.

[90]  F. Ahmad,et al.  Counteracting Osmolyte Trimethylamine N-Oxide Destabilizes Proteins at pH below Its pKa , 2005, Journal of Biological Chemistry.

[91]  S. Sligar,et al.  Changes in solvation during DNA binding and cleavage are critical to altered specificity of the EcoRI endonuclease. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[92]  T. Oas,et al.  Thermodynamic characterization of the osmolyte- and ligand-folded states of Bacillus subtilis ribonuclease P protein. , 2005, Biochemistry.

[93]  M. Record,et al.  Use of urea and glycine betaine to quantify coupled folding and probe the burial of DNA phosphates in lac repressor-lac operator binding. , 2005, Biochemistry.

[94]  K. D. Collins,et al.  The Hofmeister effect and the behaviour of water at interfaces , 1985, Quarterly Reviews of Biophysics.

[95]  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.

[96]  E. Clarke,et al.  Evaluation of the Thermodynamic Functions for Aqueous Sodium Chloride from Equilibrium and Calorimetric Measurements below 154 °C , 1985 .

[97]  Keizo Suzuki,et al.  Studies on the kinetics of protein denaturation under high pressure , 1960 .

[98]  I. Baskakov,et al.  Forcing Thermodynamically Unfolded Proteins to Fold* , 1998, The Journal of Biological Chemistry.

[99]  A. Green STUDIES IN THE PHYSICAL CHEMISTRY OF THE PROTEINS IX. THE EFFECT OF ELECTROLYTES ON THE SOLUBILITY OF HEMOGLOBIN IN SOLUTIONS OF VARYING HYDROGEN ION ACTIVITY WITH A NOTE ON THE COMPARABLE BEHAVIOR OF CASEIN , 1931 .

[100]  C. Pace,et al.  Urea and Guanidine Hydrochloride Denaturation of Ribonuclease , Lysozyme , & Zhymotrypsin , and @ Lactoglobulin * , 2003 .

[101]  D. W. Bolen,et al.  Predicting the energetics of osmolyte-induced protein folding/unfolding. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[102]  D. Rau,et al.  Preferential hydration of DNA: the magnitude and distance dependence of alcohol and polyol interactions. , 2006, Biophysical journal.

[103]  B. B. Owen,et al.  The Physical Chemistry of Electrolytic Solutions , 1963 .

[104]  G. Richmond,et al.  Isotopic Dilution Studies of the Vapor/Water Interface as Investigated by Vibrational Sum-Frequency Spectroscopy , 2002 .

[105]  C. Tanford Protein denaturation. , 1968, Advances in protein chemistry.

[106]  L. Gierasch,et al.  Inhibition of protein aggregation in vitro and in vivo by a natural osmoprotectant , 2006, Proceedings of the National Academy of Sciences.

[107]  G. Stark,et al.  Reactions of the Cyanate Present in Aqueous Urea with Amino Acids and Proteins , 1960 .

[108]  T. Kiefhaber,et al.  Kinetic coupling between protein folding and prolyl isomerization. II. Folding of ribonuclease A and ribonuclease T1. , 1992, Journal of molecular biology.

[109]  J. Gibbs On the equilibrium of heterogeneous substances , 1878, American Journal of Science and Arts.

[110]  J. Schellman,et al.  Macromolecular binding , 1975 .

[111]  T. Lohman,et al.  Thermodynamic analysis of ion effects on the binding and conformational equilibria of proteins and nucleic acids: the roles of ion association or release, screening, and ion effects on water activity , 1978, Quarterly Reviews of Biophysics.

[112]  D. G. Archer,et al.  Thermodynamic Properties of the NaCl+H2O System. II. Thermodynamic Properties of NaCl(aq), NaCl⋅2H2(cr), and Phase Equilibria , 1992 .

[113]  H. Eisenberg Protein and nucleic acid hydration and cosolvent interactions: establishment of reliable baseline values at high cosolvent concentrations. , 1994, Biophysical chemistry.

[114]  M. E. Clark,et al.  Living with water stress: evolution of osmolyte systems. , 1982, Science.

[115]  J. T. Gerig,et al.  TMAO promotes fibrillization and microtubule assembly activity in the C-terminal repeat region of tau. , 2006, Biochemistry.

[116]  A. Ben-Naim Inversion of the Kirkwood–Buff theory of solutions: Application to the water–ethanol system , 1977 .

[117]  S. Lewith,et al.  Zur Lehre von der Wirkung der Salze , 1887, Archiv für experimentelle Pathologie und Pharmakologie.

[118]  J. M. Sanchez-Ruiz,et al.  An osmolyte effect on the heat capacity change for protein folding. , 1995, Biochemistry.

[119]  B. Pettitt,et al.  Protein folding, stability, and solvation structure in osmolyte solutions. , 2005, Biophysical journal.

[120]  C. Tanford,et al.  Extension of the theory of linked functions to incorporate the effects of protein hydration. , 1969, Journal of molecular biology.

[121]  S. Shimizu,et al.  Preferential hydration and the exclusion of cosolvents from protein surfaces. , 2004, The Journal of chemical physics.

[122]  Andrew T. Russo,et al.  Osmolyte effects on kinetics of FKBP12 C22A folding coupled with prolyl isomerization. , 2003, Journal of molecular biology.

[123]  S. N. Timasheff In disperse solution, "osmotic stress" is a restricted case of preferential interactions. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[124]  Jeremy C. Smith,et al.  Protein hydration water: Structure and thermodynamics , 2002 .

[125]  D. Harries,et al.  Swelling of phospholipids by monovalent salt Published, JLR Papers in Press, November 2, 2005. , 2006, Journal of Lipid Research.

[126]  Franz Hofmeister,et al.  Zur Lehre von der Wirkung der Salze , 1888, Archiv für experimentelle Pathologie und Pharmakologie.

[127]  D. Rau,et al.  Differences in water release for the binding of EcoRI to specific and nonspecific DNA sequences. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[128]  C. Schein Solubility and secretability. , 1993, Current opinion in biotechnology.

[129]  E. L. Kovrigin,et al.  Preferential solvation changes upon lysozyme heat denaturation in mixed solvents. , 1997, Biochemistry.

[130]  H. Viadiu,et al.  Structure of BamHI bound to nonspecific DNA: a model for DNA sliding. , 2000, Molecular cell.

[131]  Adam W Van Wynsberghe,et al.  The exclusion of glycine betaine from anionic biopolymer surface: why glycine betaine is an effective osmoprotectant but also a compatible solute. , 2004, Biochemistry.

[132]  C. Pace,et al.  Denaturant m values and heat capacity changes: Relation to changes in accessible surface areas of protein unfolding , 1995, Protein science : a publication of the Protein Society.

[133]  C. Tanford,et al.  Proteins as random coils. II. Hydrogen ion titration curve of ribonuclease in 6 M guanidine hydrochloride. , 1967, Journal of the American Chemical Society.

[134]  E. Galinski,et al.  Osmoadaptation in bacteria. , 1995, Advances in microbial physiology.

[135]  W. Neuhofer,et al.  Cell survival in the hostile environment of the renal medulla. , 2005, Annual review of physiology.

[136]  M. Kozlov,et al.  Bending, hydration and interstitial energies quantitatively account for the hexagonal-lamellar-hexagonal reentrant phase transition in dioleoylphosphatidylethanolamine. , 1994, Biophysical journal.

[137]  A. Rosengarth,et al.  Slow unfolding and refolding kinetics of the mesophilic Rop wild-type protein in the transition range. , 1999, European journal of biochemistry.

[138]  V. Parsegian,et al.  Protein solvation in allosteric regulation: a water effect on hemoglobin. , 1992, Science.

[139]  D. W. Bolen,et al.  Mixed osmolytes: The degree to which one osmolyte affects the protein stabilizing ability of another , 2006, Protein science : a publication of the Protein Society.

[140]  M. R. Sheen,et al.  How tonicity regulates genes: story of TonEBP transcriptional activator , 2006, Acta physiologica.

[141]  Arieh Ben-Naim,et al.  Statistical Thermodynamics for Chemists and Biochemists , 1992, Springer US.

[142]  M. Yao,et al.  How valid are denaturant-induced unfolding free energy measurements? Level of conformance to common assumptions over an extended range of ribonuclease A stability. , 1995, Biochemistry.

[143]  M G Fried,et al.  Role of macromolecular hydration in the binding of the Escherichia coli cyclic AMP receptor to DNA. , 1997, Biochemistry.

[144]  The effect of ion pairs on the thermal stability of D-glyceraldehyde 3-phosphate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima. , 1994, Protein engineering.

[145]  S. Hawley,et al.  Reversible pressure--temperature denaturation of chymotrypsinogen. , 1971, Biochemistry.

[146]  Ashutosh Tripathy,et al.  Effects of molecular crowding by saccharides on alpha-chymotrypsin dimerization. , 2002, Protein science : a publication of the Protein Society.

[147]  V. Adrian Parsegian,et al.  Polymer inaccessible volume changes during opening and closing of a voltage-dependent ionic channel , 1986, Nature.

[148]  S. Bezrukov,et al.  Probing alamethicin channels with water-soluble polymers. Effect on conductance of channel states. , 1993, Biophysical journal.

[149]  G. Rose,et al.  Modeling unfolded states of proteins and peptides. II. Backbone solvent accessibility. , 1997, Biochemistry.

[150]  S. Weerasinghe,et al.  A Kirkwood-Buff derived force field for the simulation of aqueous guanidinium chloride solutions. , 2004, The Journal of chemical physics.

[151]  V A Parsegian,et al.  Osmotic stress, crowding, preferential hydration, and binding: A comparison of perspectives. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[152]  Charles Tanford,et al.  Isothermal Unfolding of Globular Proteins in Aqueous Urea Solutions , 1964 .

[153]  D. Rau,et al.  Water release associated with specific binding of gal repressor. , 1995, The EMBO journal.

[154]  K. Yutani,et al.  The unusually slow unfolding rate causes the high stability of pyrrolidone carboxyl peptidase from a hyperthermophile, Pyrococcus furiosus: equilibrium and kinetic studies of guanidine hydrochloride-induced unfolding and refolding. , 1998, Biochemistry.

[155]  S. K. Woo,et al.  Tonicity-responsive enhancer binding protein, a rel-like protein that stimulates transcription in response to hypertonicity. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[156]  I. Baskakov,et al.  The osmophobic effect: natural selection of a thermodynamic force in protein folding. , 2001, Journal of molecular biology.

[157]  R. Mazo,et al.  On the Statistical Mechanical Theory of Solutions , 1958 .

[158]  K. Tsumoto,et al.  The effects of arginine on refolding of aggregated proteins: not facilitate refolding, but suppress aggregation. , 2003, Biochemical and biophysical research communications.

[159]  Paul E Smith,et al.  Protein volume changes on cosolvent denaturation. , 2005, Biophysical chemistry.

[160]  G. Pielak,et al.  Impact of protein denaturants and stabilizers on water structure. , 2004, Journal of the American Chemical Society.

[161]  L. Serrano,et al.  NMR and SAXS characterization of the denatured state of the chemotactic protein Che Y: Implications for protein folding initiation , 2001, Protein science : a publication of the Protein Society.

[162]  C. Pace,et al.  Buried, charged, non-ion-paired aspartic acid 76 contributes favorably to the conformational stability of ribonuclease T1. , 1999, Biochemistry.

[163]  C. Pace,et al.  Urea and guanidine hydrochloride denaturation of ribonuclease, lysozyme, alpha-chymotrypsin, and beta-lactoglobulin. , 1974, The Journal of biological chemistry.

[164]  D. Rau,et al.  Differences between EcoRI nonspecific and "star" sequence complexes revealed by osmotic stress. , 2004, Biophysical journal.

[165]  Kevin L. Shaw,et al.  The effect of net charge on the solubility, activity, and stability of ribonuclease Sa , 2001, Protein science : a publication of the Protein Society.

[166]  Seishi Shimizu,et al.  Preferential hydration of proteins: A Kirkwood-Buff approach , 2006 .

[167]  Y. Shamoo,et al.  Photochemical crosslinking of bacteriophage T4 single‐stranded DNA‐binding protein (gp32) to oligo‐p(dT)8: Identification of phenylalanine‐183 as the site of crosslinking , 1988, Proteins.

[168]  E. Ruckenstein,et al.  Relationship between preferential interaction of a protein in an aqueous mixed solvent and its solubility. , 2005, Biophysical chemistry.

[169]  Paul E. Smith,et al.  Cosolvent Interactions with Biomolecules: Relating Computer Simulation Data to Experimental Thermodynamic Data , 2004 .

[170]  A. Green STUDIES IN THE PHYSICAL CHEMISTRY OF THE PROTEINS IX. THE EFFECT OF ELECTROLYTES ON THE SOLUBILITY OF HEMOGLOBIN IN SOLUTIONS OF VARYING HYDROGEN ION ACTIVITY WITH A NOTE ON THE COMPARABLE BEHAVIOR OF CASEIN , 1931 .

[171]  S. Weerasinghe,et al.  A Kirkwood−Buff Derived Force Field for Mixtures of Urea and Water , 2003 .

[172]  H. Hinz,et al.  Pressure-modulated differential scanning calorimetry. An approach to the continuous, simultaneous determination of heat capacities and expansion coefficients. , 2006, Analytical chemistry.

[173]  R. DesJarlais,et al.  Effect of construct design on MAPKAP kinase-2 activity, thermodynamic stability and ligand-binding affinity. , 2006, Archives of biochemistry and biophysics.

[174]  V A Parsegian,et al.  Probing alamethicin channels with water-soluble polymers. Size-modulated osmotic action. , 1993, Biophysical journal.

[175]  S. Shimizu Estimating hydration changes upon biomolecular reactions from osmotic stress, high pressure, and preferential hydration experiments. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[176]  D. W. Bolen,et al.  Additive transfer free energies of the peptide backbone unit that are independent of the model compound and the choice of concentration scale. , 2004, Biochemistry.

[177]  E. Eisenstein,et al.  The effect of water activity on the association constant and the enthalpy of reaction between lysozyme and the specific antibodies D1.3 and D44.1 , 1996, Journal of molecular recognition : JMR.

[178]  K. Heremans,et al.  High pressure effects on proteins and other biomolecules. , 1982, Annual review of biophysics and bioengineering.

[179]  A. Aggarwal,et al.  Structure of Bam HI endonuclease bound to DNA: partial folding and unfolding on DNA binding. , 1995, Science.

[180]  G. Somero,et al.  Biochemical Adaptation: Mechanism and Process in Physiological Evolution , 1984 .

[181]  G. Bomhoff,et al.  The effects of the flavonoid baicalein and osmolytes on the Mg 2+ accelerated aggregation/fibrillation of carboxymethylated bovine 1SS-alpha-lactalbumin. , 2006, Archives of biochemistry and biophysics.

[182]  Robin S. Dothager,et al.  Random-coil behavior and the dimensions of chemically unfolded proteins. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[183]  V Adrian Parsegian,et al.  Protein-water interactions. , 2002, International review of cytology.

[184]  C. Tanford,et al.  Theory of Protein Titration Curves. I. General Equations for Impenetrable Spheres , 1957 .

[185]  Lars Onsager,et al.  The Surface Tension of Debye‐Hückel Electrolytes , 1934 .

[186]  N. Templeton,et al.  Gene therapy : therapeutic mechanisms and strategies , 2000 .

[187]  Seishi Shimizu,et al.  The Kirkwood-Buff theory and the effect of cosolvents on biochemical reactions. , 2004, The Journal of chemical physics.

[188]  J. Kirkwood,et al.  The Statistical Mechanical Theory of Solutions. I , 1951 .

[189]  M. Record,et al.  Application of the local-bulk partitioning and competitive binding models to interpret preferential interactions of glycine betaine and urea with protein surface. , 2004, Biochemistry.

[190]  T. Chalikian Structural Thermodynamics of Hydration , 2001 .

[191]  V. Parsegian,et al.  Macromolecules and water: probing with osmotic stress. , 1995, Methods in enzymology.

[192]  E. Cohn,et al.  STUDIES IN THE PHYSICAL CHEMISTRY OF THE PROTEINS , 1924, The Journal of general physiology.

[193]  G. Pielak,et al.  Effects of crowding by mono-, di-, and tetrasaccharides on cytochrome c-cytochrome c peroxidase binding: comparing experiment to theory. , 2001, Biochemistry.

[194]  G. Richmond,et al.  Isolated Molecular Ion Solvation at an Oil/Water Interface Investigated by Vibrational Sum-Frequency Spectroscopy , 2004 .

[195]  W. Emmerich,et al.  Effect of polyethylene glycol exclusion on the water potential of solution-saturated filter paper. , 1990, Plant physiology.

[196]  D. Harries,et al.  Solutes probe hydration in specific association of cyclodextrin and adamantane. , 2005, Journal of the American Chemical Society.

[197]  R. Santucci,et al.  Influence of glycerol on the structure and stability of ferric horse heart myoglobin: a SAXS and circular dichroism study. , 1996, Biochimica et biophysica acta.

[198]  K. Spiro Ueber die Beeinflussung der Eiweisscoagulation durch stickstoffhaltige Substanzen. , 1900 .

[199]  Doug Barrick,et al.  Measuring the stability of partly folded proteins using TMAO , 2003, Protein science : a publication of the Protein Society.

[200]  D. W. Bolen,et al.  Unfolding free energy changes determined by the linear extrapolation method. 1. Unfolding of phenylmethanesulfonyl alpha-chymotrypsin using different denaturants. , 1988, Biochemistry.

[201]  S. Krimm,et al.  Extended conformations of polypeptides and proteins in urea and guanidine hydrochloride , 1973 .

[202]  B. E. Davidson,et al.  Effects of molecular crowding on the interaction between DNA and the Escherichia coli regulatory protein TyrR. , 1997, Biophysical journal.

[203]  V A Parsegian,et al.  Parametrization of direct and soft steric-undulatory forces between DNA double helical polyelectrolytes in solutions of several different anions and cations. , 1994, Biophysical journal.

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

[205]  P. Yancey,et al.  Organic osmolytes as compatible, metabolic and counteracting cytoprotectants in high osmolarity and other stresses , 2005, Journal of Experimental Biology.

[206]  G. Conn,et al.  Ion chromatographic quantification of cyanate in urea solutions: estimation of the efficiency of cyanate scavengers for use in recombinant protein manufacturing. , 2004, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[207]  S. Leikin,et al.  Interactions of inorganic phosphate and sulfate anions with collagen. , 2004, Biochemistry.

[208]  C. Russell Middaugh,et al.  Formulation Design of Acidic Fibroblast Growth Factor , 1993, Pharmaceutical Research.

[209]  K. D. Collins Ion hydration: Implications for cellular function, polyelectrolytes, and protein crystallization. , 2006, Biophysical chemistry.

[210]  C. Pace,et al.  Protein structure, stability and solubility in water and other solvents. , 2004, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[211]  S. L. Sörensen THE SOLUBILITY OF PROTEINS , 1925 .

[212]  H. Hinz,et al.  Phase diagrams: a graphical representation of linkage relations. , 2003, Journal of molecular biology.

[213]  J. Kornblatt,et al.  The effects of osmotic and hydrostatic pressures on macromolecular systems. , 2002, Biochimica et Biophysica Acta.

[214]  Hoang T. Tran,et al.  Toward an accurate theoretical framework for describing ensembles for proteins under strongly denaturing conditions. , 2006, Biophysical journal.

[215]  J. Lee,et al.  The Conformation of the Glucocorticoid Receptor AF1/tau1 Domain Induced by Osmolyte Binds Co-regulatory Proteins* , 2001, The Journal of Biological Chemistry.

[216]  K. Bailey Proteins, Amino-Acids and Peptides as Ions and Dipolar Ions , 1945, Nature.

[217]  Philip Yeagle,et al.  The Structure of Biological Membranes , 2004 .

[218]  H. Hinz,et al.  Pressure-modulated differential scanning calorimetry: theoretical background. , 2006, Analytical chemistry.

[219]  J. Wyman,et al.  LINKED FUNCTIONS AND RECIPROCAL EFFECTS IN HEMOGLOBIN: A SECOND LOOK. , 1964, Advances in protein chemistry.

[220]  M. Fried,et al.  Role of Hydration in the Binding of lac Repressor to DNA* , 2002, The Journal of Biological Chemistry.

[221]  C. F. Anderson,et al.  Thermodynamic Expressions Relating Different Types of Preferential Interaction Coefficients in Solutions Containing Two Solute Components , 2002 .

[222]  H. Hinz,et al.  A new alternative method to quantify residual structure in 'unfolded' proteins. , 2000, Biochimica et biophysica acta.

[223]  J. Michael Schurr,et al.  A contribution to the theory of preferential interaction coefficients. , 2005, Biophysical journal.

[224]  Paul E. Smith,et al.  Equilibrium dialysis data and the relationships between preferential interaction parameters for biological systems in terms of Kirkwood-Buff integrals. , 2006, The journal of physical chemistry. B.