Biophysical Reviews

, Volume 10, Issue 2, pp 203–208 | Cite as

Protein–solvent interaction

  • Tsutomu ArakawaEmail author


Protein folding and assembly can be manipulated in in vitro systems by co-solvents at high concentrations. A number of co-solvents that enhance protein stability and assembly have been shown to be excluded from the protein surface. Such co-solvent exclusion has been demonstrated by dialysis experiments and shown to be correlated with their effects on protein stability and assembly.


Protein folding and assembly In vitro systems Co-solvents Preferentially excluded co-solvents Dialysis equilibrium 


Compliance with ethical standards

Conflict of interest

Tsutomu Arakawa declares that he has no conflicts of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by the author.


  1. Algaier J, Himes RH (1989) The effects of dimethyl sulfoxide on the kinetics of tubulin assembly. Biochim Biophys Acta 954:235–243CrossRefGoogle Scholar
  2. Arakawa T, Timasheff SN (1982a) Preferential interactions of proteins with salts in concentrated solutions. Biochemistry 21:6545–6552CrossRefPubMedGoogle Scholar
  3. Arakawa T, Timasheff SN (1982b) Stabilization of protein structure by sugars. Biochemistry 21:6536–6544CrossRefPubMedGoogle Scholar
  4. Arakawa T, Timasheff SN (1985a) Mechanism of poly(ethylene glycol) interaction with proteins. Biochemistry 24:6756–6762CrossRefPubMedGoogle Scholar
  5. Arakawa T, Timasheff SN (1985b) The stabilization of proteins by osmolytes. Biophys J 47:411–414CrossRefPubMedPubMedCentralGoogle Scholar
  6. Arakawa T, Carpenter JF, Kita Y, Crowe JH (1990) The basis for toxicity of cartain cryoprotectants: a hypothesis. Cryobiology 27:401–415CrossRefGoogle Scholar
  7. Arakawa T, Kita Y, Carpenter JF (1991) Protein-solvent interactions in pharmaceutical formulations. Pharm Res 8:285–291CrossRefPubMedGoogle Scholar
  8. Asakura S, Oosaw F (1954) On interaction beween two bodies immersed in a solution of macromolecules. J Phys Chem 22:1255–1256CrossRefGoogle Scholar
  9. Carpenter JF, Crowe JH (1988) The mechanism of cryoprotection of proteins by solutes. Cryobiology 25:244–255CrossRefPubMedGoogle Scholar
  10. De La Cruz EM, Pollard TO (1995) Nucleotide-free actin: stabilization by sucrose and nucleotide binding kinetics. Biochemistry 34:5452–5461CrossRefGoogle Scholar
  11. Durham AC (1972) Structure and roles of the polymorphic forms of tobacco mosaic virus protein. I Sedimentation studies. J Mol Biol 67:289–305CrossRefPubMedGoogle Scholar
  12. Erickson HP (1974a) Assembly of microtubules from preformed, ring-shaped protofilaments and 6-S tubulin. J Supramol Struct 2:393–411CrossRefPubMedGoogle Scholar
  13. Erickson HP (1974b) Microtubule surface lattice and subunit structure and observations on reassembly. J Cell Biol 60:153–167CrossRefPubMedPubMedCentralGoogle Scholar
  14. Gagnon P (2008) Improved antibody aggregate removal by hydroxyapatite chromatography in the presence of polyethylene glycol. J Immunol Methods 336:222–228CrossRefPubMedGoogle Scholar
  15. Gagnon P, Toh P, Lee J (2014) High productivity purification of immunoglobulin G monoclonal antibodies on starch-coated magnetic nanoparticles by steric exclusion of polyethylene glycol. J Chromatogr A 1324:171–180CrossRefPubMedGoogle Scholar
  16. Gekko K, Timasheff SN (1981) Mechanism of protein stabilization by glycerol: preferential hydration in glycerol–water mixtures. Biochemistry 20:4667–4676Google Scholar
  17. Gibbs JW (1878) On the equilibrium of heterogeneous substances. Trans Connecticut Acad Arts Sci 3:343–524Google Scholar
  18. Hamel E, Lin CM (1981a) Glutamate-induced polymerization of tubulin: characterization of the reaction and application to the large-scale purification of tubulin. Arch Biochem Biophys 209:29–40CrossRefPubMedGoogle Scholar
  19. Hamel E, Lin CM (1981b) Interaction of tubulin with ribose-modified analogs of GTP and GDP: evidence for two mutually exclusive exchangeable nucleotide binding sites. Proc Natl Acad Sci USA 78:3368–3372CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hamel E, del Campo AA, Lowe MC, Lin CM (1981) Interaction of taxol, microtubule-associated proteins, and guanine nucleotides in tubulin polymerization. J Biol Chem 256:11887–11894PubMedGoogle Scholar
  21. Hamel E, del Campo AA, Lowe MC, Waxman PG, Lin MC (1982) Effects of organic acids on tubulin polymerization and associated guanosine 5′-triphosphate hydrolysis. Biochemistry 21:503–509CrossRefPubMedGoogle Scholar
  22. Iqbal K, Liu F, Gong CX, Grunde-Iqbal I (2010) Tau in Alzheimer diseases and related tauopathies. Curr Alzheimer Res 7:656–664CrossRefPubMedPubMedCentralGoogle Scholar
  23. Kasai M (1969) Thermodynamical aspect of G-F transformations of actin. Biochim Biophys Acta 180:399–409CrossRefPubMedGoogle Scholar
  24. Kasai M, Nakano E, Oosawa F (1965) Polymerization of actin free from nucleotides and divalent cations. Biochim Biophys Acta 29:494–503CrossRefGoogle Scholar
  25. Klug A (1999) The tobacco mosaic virus particles: structure ad assembly. Philos Trans R Soc Lond B 354:531–535CrossRefGoogle Scholar
  26. Lauffer MA, Stevens CL (1968) Structure of the tobacco mosaic virus particles; polymerization of tobacco mosaic virus protein. Adv Virus Res 13:1–63CrossRefPubMedGoogle Scholar
  27. Lee JC, Lee LL (1979) Interaction of calf brain tubulin with poly(ethylene glycols). Biochemistry 18:5518–5526CrossRefPubMedGoogle Scholar
  28. Lee JC, Timasheff SN (1977) In vitro reconstitution of calf brain microtubules: effects of solution variables. Biochemistry 19:1754–1764CrossRefGoogle Scholar
  29. Lee JC, Timasheff SN (1981) The stabilization of proteins by sucrose. J Biol Chem 256:7193–7201PubMedGoogle Scholar
  30. Lee JC, Gekko K, Timasheff SN (1979) Measurement of preferential solvent interactions by densimetric techniques. Methods Enzymol 61:26–49CrossRefPubMedGoogle Scholar
  31. Lee J, Gan HT, Latiff SM, Chuah C, Lee WY, Yang YS, Loo B, Ng SK, Gagnon P (2012) Principles and application of steric exclusion chromatography. J Chromatogr A 1270:162–170CrossRefPubMedGoogle Scholar
  32. Mackenzie KF, Singh KK, Brown AD (1988) Water stress plating hypersensitivity of yeasts: protective role of trehalose in Saccharomyces Cerevisiae. J Gen Microbiol 134:1661–1666PubMedGoogle Scholar
  33. Melander W, Horvath C (1977) Salt effect on hydrophobic interactions in precipitation and chromatography of proteins: an interpretation of the lyotropic series. Arch Biochem Biophys 183:200–215CrossRefPubMedGoogle Scholar
  34. Minton AP (1980) Excluded volume as a determinant of protein structure and stability. Biophys J 32:77–79CrossRefPubMedPubMedCentralGoogle Scholar
  35. Na GC, Timasheff SN (1981) Interaction of calf brain tubulin with glycerol. J Mol Biol 151:165–178CrossRefPubMedGoogle Scholar
  36. Nagy B, Jencks WP (1965) Depolymerization of F-actin by concentrated solutions of salts and denaturing agents. J Am Chem Soc 87:2480–2488CrossRefPubMedGoogle Scholar
  37. Oosawa F, Kasai M (1962) A theory of linear and helical aggregations of macromolecules. J Mol Biol 4:10–21CrossRefPubMedGoogle Scholar
  38. Pittz EP, Timasheff SN (1978) Interaction of ribonuclease a with 2-methyl-2,4-pentanediol at pH 5.8. Biochemistry 17:615–623CrossRefPubMedGoogle Scholar
  39. Ramkumar A, Jong BY, Ori-McKenney KM (2017) ReMAPpng the microtubule landscape: how phosphorylation dictates the activities of microtubule-associated proteins. Dev Dyn.
  40. Rebhun LI, Sawada N (1969) Augmentation and dispersion of the in vivo mitotic apparatus of living marine eggs. Protoplasma 68:1–22CrossRefPubMedGoogle Scholar
  41. Rebhun LI, Mellon M, Jemiolo D, Nath J, Ivy N (1974) Regulation of size and birefringence of the in vivo apparatus. J Supramol Struct 2:466–485CrossRefPubMedGoogle Scholar
  42. Sánchez MP, Alvarez-Tallada V, Avila J (2001) The microtubule-associated protein tau in neurodegenerative diseases. Tauropathies. Rev Neurol 33:169–177PubMedGoogle Scholar
  43. Schellman JA (2003) Protein stability in mixed solvents: a balance of contact interaction and excluded volume. Biophys J 85:108–125CrossRefPubMedPubMedCentralGoogle Scholar
  44. Sheawin KE, Winzor DJ (1988) Effect of sucrose on the dimerization of alpha-chymotrypsin. Allowance for thermodynamic nonideality arising from the presence of aa small inert solute. Biophys Chem 31:287–294CrossRefGoogle Scholar
  45. Shelanski ML, Gaskin F, Cantor CR (1973) Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci USA 70:765–768CrossRefPubMedPubMedCentralGoogle Scholar
  46. Sheterline P, Schofield JG, Mira-Moser F (1977) The effect of secretotogues and 2-methylpentane-2,4-diol on the microtubule-tubulin equilibrium and the release of growth hormone from bovine anterior pituitary slices. Exp Cell Res 104:127–134CrossRefPubMedGoogle Scholar
  47. Sloboda RD, Dentler WL, Rosenbaum JL (1976) Microtubule-associated proteins and the stimulation of tubulin assembly in vitro. Biochemistry 15:4497–4505CrossRefPubMedGoogle Scholar
  48. Snyder MA, Ng P, Mekosh H, Gagnon P (2009) PEG enhances viral clearance on ceramic hydroxyapatite. J Sep Sci 32:4048–4051CrossRefPubMedGoogle Scholar
  49. Timasheff SN (1992) Water as ligand: preferential binding and exclusion of denaturants in protein unfolding. Biochemistry 31:9857–9864CrossRefPubMedGoogle Scholar
  50. Timasheff SN (1993) The control of protein stability and association by weak interactions with water: how do solvents affect these processes. Annu Rev Biophys Biomol Struct 22:67–97CrossRefPubMedGoogle Scholar
  51. Timasheff SN (1998) Control of protein stability and reactions by weakly interacting cosolvents: the simplicity of the complicated. Adv Protein Chem 51:355–432CrossRefPubMedGoogle Scholar
  52. Timasheff SN (2002) Thermodynamic binding and site occupancy in the light of the Schellman exchange concept. Biophys Chem 102-102:99–111CrossRefGoogle Scholar
  53. Timasheff SN, Inoue H (1968) Preferential binding of solvent components to proteins in mixed water–organic solvent systems. Biochemistry 7:2501–2513Google Scholar
  54. Traube J (1910) The attraction pressure. J Phys Chem 14:451–470Google Scholar
  55. Wakabayashi K, Hotani H, Asakura S (1969) Polymerization of salmonella flagellin in the presence of high concentrations of salts. Biochim Biophys Acta 175:195–203CrossRefPubMedGoogle Scholar
  56. Yancey PH, Clark ME, Hand SC, Bowlus RD, Somero GN (1982) Living with water stress: evolution of osmolyte systems. Science 217:1214–1222CrossRefPubMedGoogle Scholar
  57. Yoshimoto N, Itoh D, Isakari Y, Podgomik A, Yamamoto S (2015) Salt tolerant chromatography provides salt tolerance and a better selectivity for protein monomer separations. Biotechnol J 10:1929–1934CrossRefPubMedGoogle Scholar
  58. Zhang W, Capp MW, Bond JP, Anderson CF, Record MT Jr (1996) Thermodynamic characterization of interactions of native bovine serum albumin with highly excluded (glycine betaine) and moderately accumulated (urea) solutes by a novel application of vaport pressure osmometry. Biochemistry 35:10506–10516Google Scholar

Copyright information

© International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Alliance Protein Laboratories, a Division of KBI BiopharmaSan DiegoUSA

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