Abstract
The study of pressure effects on protein stability has occupied a relatively marginal position in the field of protein folding, with very few thorough thermodynamic, structural and kinetic studies of this phenomenon. Moreover, theoretical treatment of the issue with a few recent exceptions, has been limited to declarations of its complexity and lack of concordance with the results from other approaches. This paucity of data and theory notwithstanding, understanding the fundamental physical basis for pressure effects on proteins is essential to progress in the field of protein folding. Moreover, pressure presents certain advantages as a perturbation methodology that render it an important, useful and complementary approach. In the present review, the issue of the fundamental basis for the effects of pressure is discussed. Reference is made to studies in the literature, but I have concentrated the detailed presentation on the body of work in pressure-induced protein unfolding carried out by my research group and collaborators on staphylococcal nuclease (Snase) over the past 5 years. The origins of the value of the change in volume upon unfolding must be understood prior to any thorough theoretical analysis of pressure effects. The various arguments for the multiple contributing factors are discussed and then recent studies from my research group designed to probe this question are presented, the overall conclusion being that the existence of packing defects in the folded structure represents the most likely candidate for the negative change in volume upon unfolding. Moreover, the results of the temperature dependence of the volume change for unfolding of Snase implicate the difference in thermal expansivity in the temperature dependence of the value of the volume change of unfolding. Next I present results of a characterization of the physical properties of the pressure denatured state of Snase, and compare these to studies on a number of other pressure denatured proteins. Finally, the results of a series of pressure-jump kinetic studies on the folding/unfolding reactions of this protein are discussed. It is too early to conclude whether the results from these pressure studies on Snase stability and their interpretations are general. For this, many more studies on a number of small, reversibly folding proteins will be required.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
1. Kauzmann, W (1959) Some factors in the interpretation of protein denaturation in Advances in Protein Chemistry in J C B Anfinsen, K Bailey, M L Anson, and J T Edsall, eds. Academic Press, New York and London, 1–66.
2. Kim, P. S. & Baldwin, R. L. (1990) Intermediates in the folding reactions of small proteins, Annu. Rev. Biochem. 59, 631–660.
3. Onuchic, J. N., Luthey-Schulten, Z. & Wolynes, P. G. (1997) Theory of protein folding: the energy landscape perspective, Annu. Rev. Phys. Chem. 48, 545–600.
4. Onuchic, J. N., Wolynes, P. G., Luthey-Schulten, Z. & Socci, N. D. (1995) Toward an outline of the topography of a realistic protein folding funnel Proc. Natl. Acad. Sci. USA 92, 3626–3630.
5. Socci, N. D., Onuchic, J. N. & Wolynes, P. G. (1996) Diffusive dynamics for the reaction coordinate for protein folding funnels, J. Chem. Phys. 104, 5860–5868.
6. Privalov, P L & Gill, S J (1988) Stability of protein structure and hydrophobic interaction, Adv. Prot. Chem. 39, 191–234.
7. Brandts, J F, Oliveira, R J & Westort, C (1970) Thermodynamics of protein denaturation: Effects of pressure on the denaturation of ribonuclease A, Biochemistry, 9, 1038–104
8. Zipp, A & Kauzmann, W (1973) Pressure denaturation of metmyoglobin, Biochemistry, 12, 4217–422
9. Katz, S, Crissman, J K Jr. & Beall, J A (1973) Structure volume relationships of proteins: Dilatometric studies of the structural transitions engendered in serum albumin and myoglobin as a consequence of acid-base reaction in water and in denaturing media, J. Biol. Chem. 248, 4840–4845.
10. Suzuki, K, Miyosawa, Y & Suzuki, C (1963) Protein denaturation by high pressure. Measurements of turbidity of isoelectric ovalbumin and horse serum albumin under high pressure, Arch. Biochem. Biophys., 101, 225–22
11. Li, T M, Hook, T W, Drickamer, H G & Weber, G (1976) Plurality of pressure denatured forms in chymotrypsinogen and lysozyme, Biochemistry, 15, 5572–558
12. Samarasinghe, S D, Campbell, D M, Jonas, A & Jonas, J (1992) High resolution NMR study of the pressure-induced unfolding of lysozyme, Biochemistry, 31, 7773–777
13. Hawley, S A (1971) Reversible temperature-pressure denaturation of chymotrypsinogen, Biochemistry, 10, 2436–244
14. Wong, P T T & Heremans, K (1988) Pressure effects on protein secondary structure and hydrogen deuterium exchange in chymotrypsinogen: A Fourier transform infrared spectroscopy study, Biochim. Biophys. Acta., 965, 1–9
15. Marden, M C, Hui Bon Hoa, G & Marden-Stetzkowski, F (1986) Heme protein fluorescence vs. pressure Biophys. J. 49, 619–627.
16. Takeda, N, Kato, M & Taniguchi, Y (1995) Pressure and thermally induced reversible changes in the secondary structure of ribonuclease A, Biochemistry, 34, 5980–598
17. Zhang, J., Peng, X., Jonas, A. & Jonas, J. (1995) NMR study of the cold, heat and pressure unfolding of ribonuclease A, Biochemistry 34, 8631–8641.
18. Nash D., Lee B.S., & Jonas J. (1996) Hydrogen exchange kinetics in the cold denatured state of ribonuclease A, Biochim. Biophys. Acta, 1297, 40–4
19. Royer, C A, Hinck, A P, Loh, S N, Prehoda, K E, Peng, X, Jonas, J & Markley, J L (1993) Effects of amino acid substitutions on the pressure denaturation of staphylococcal nuclease as monitored by fluorescence and nuclear magnetic resonance spectroscopy, Biochemistry, 32, 5222–523
20. Vidugiris, G J A, Markley, J L & Royer, C A (1995) Evidence for a molten globule-like transition state in protein folding, Biochemistry, 34, 4909–491
21. Vidugiris, G J A, Truckses, D M, Markley, J L & Royer, C A (1996) High pressure denaturation of staphylococcal nuclease proline to glycine substitution mutants, Biochemistry, 35, 3857–386
22. Frye, K J, Perman, C S & Royer, C A (1996) Testing the correlation between ΓA and ΓV in protein unfolding using m-value mutants of staphylococcal nuclease, Biochemistry, 35, 10234–1023
23. Frye, K J & Royer, C A (1997) The kinetic basis for the stabilization of staphylococcal nuclease by xylose, Prot. Sci., 6, 789–79
24. Eftink, M R, Ghiron, C A, Kautz, R A, & Fox, R O (1991) Fluorescence studies with staphylococcal nuclease and its site directed mutant, Biochemistry, 30, 1193–119
25. Eftink, M R & Ramsay, G D (1996) Temperature and pressure induced unfolding of a mutant of staphylococcal nuclease A, In High-Pressure Effects in Molecular Biophysics and Enzymology. J. L. Markley, D. B. Northrop, and C. A. Royer, editors. Oxford University Press, New York. 62–73.
26. Dill, K A (1990)Dominant forces in protein folding, Biochemistry 29, 7133–7155.
27. Mozhaev, V V, Heremans, K, Frank, J, Masson, P & Balny, C (1996) High pressure effects on protein structure and function, Proteins: Struct. Funct. Genet., 24, 81–9
28. Kauzmann, W. (1986) Protein stabilization: Thermodynamics of unfolding, Nature 325, 763–764.
29. Klapper, M. H. (1971) On the nature of the protein interior, Biochim. Biophys. Acta 229, 557–566.
30. Hvidt, A. (1975) A discussion of pressure-volume effects in aqueous protein solutions, J. theor. Biol. 50, 245–252.
31. Masterton, W L (1954) Partial molar volumes of hydrocarbons in solution, J. Chem. Phys. 22, 1830–1833.
32. Assarsson, P & Eirich, F R (1968) Properties of amides in aqueous solution. I. A. Viscosity and density changes of amide-water systems. B. An analysis of volume deficiencies of mixtures based on molecular size differences (Mixing of hard spheres), J. Phys. Chem. 72, 2710–2719.
33. Prehoda, K E, & Markley, J L (1996) Use of partial molar volumes of model compounds in the interpretation of high pressure effects on proteins, In High-Pressure Effects in Molecular Biophysics and Enzymology. J L Markley, D B Northrop, and C A. Royer, eds. Oxford University Press, New York. 33–43.
34. Frank, H. S. & Evans, M. W. (1945) Free volume and entropy in condensed systems: III Entropy in binary liquid mixtures, partial molal entropy in dilute solutions, structure and thermodynamics of aqueous electrolytes, J. Phys. Chem. 13, 507–532.
35. Masterton, W L & Seiler, H (1968) Apparent and partial molar volumes of water in organic solvents, J. Phys. Chem. 72, 4257–4262.
36. Richards, F M (1977) Areas, volumes, packing and protein structure, Ann. Rev. Biophys. Bioeng. 6, 151–176.
37. Rashin A A Iofin M & Honig B (1986) Internal cavities and buried waters in globular proteins Biochemistry 25: 3619–3625
38. Nogochi, H & Yang, JT (1963) Dilatometric and refractometric studies of the helix-coil transition of poly-glutamic acid in aqueous solutions, Biopolymers 1, 359–370.
39. Weber, G & Drickamer, H G (1983) The effect of high pressure on proteins and other biomolecules, Quart Rev. Biophys., 16, 89–11
40. Jaenicke, R (1981) Enzymes under extreme conditions, Ann. Rev. Biophys. Bioeng. 10, 1–67.
41. Heremans, K (1982) High pressure effects on proteins and other biomolecules, Annu. Rev Biophy. Bioeng. 11, 1–21.
42. Chalikian, T V and Breslauer K J (1996) On volume changes accompanying conformational transitions of biopolymers, Biopolymers 39, 619–626.
43. Hummer, G., Garde, S., Garcia, A.E. Paulaitis, ME. & Pratt, L. R. (1998) The pressure dependence of hydrophobic interactions is consistent with the observed pressure denaturation of proteins, Proc. Natl. Acad. Sci. USA 95, 1552–1555.
44. Hummer, G., Garde, S., Garcia, A. E., Pohorille, A. & Pratt, L. R. (1996) An information theory model of hydropobic interactions, Proc. Natl. Acad. Sci. USA 93, 8951–8955.
45. Murphy, KP & Freire, E. (1992) Thermodynamics of structural stability and cooperative folding behavior in proteins, Adv. Prot. Chem. 43, 313–361.
46. Hynes, T R & Fox, R O (1991) The crystal structure of staphylococcal nuclease refines at 1.7 Å resolution, Proteins: Struct. Funct. Genet., 10, 92–10
47. Shortle, D & Meeker, A K (1986) Mutant forms of staphylococcal nuclease with altered patterns of guanidine hydrochloride and urea denaturation, Proteins: Struct. Funct. Genet., 1, 81–8
48. Carra, J H & Privalov, P L (1995)Energetics of denaturation and m-values of staphylococcal nuclease mutants, Biochemistry, 34, 2034–204
49. Chen, H M, You, J L, Markin, V S & Tsong, T Y (1991) Kinetic analysis of the acid and alkaline unfolded states of staphylococcal nuclease, J. Mol. Biol. 220, 771–778.
50. Chen, H M, Markin, V S & Tsong, T Y (1992) pH-induced folding/unfolding of staphylococcal nuclease: Determination of kinetic parameters by the sequential jump method, Biochemistry, 31, 1483–149
51. Su Z D Arooz M T Chen H M Gross C J & Tsong T Y (1996) Least activation path for protein folding: investigation of staphylococcal nuclease folding by stopped-flow circular dichroism Proc. Natl. Acad. Sci. USA 93 2539–2544
52. Shortle, D, Meeker, A K & Gerring, S L (1989) Effects of denaturant at low concentrations on the reversible denaturation of staphylococcal nuclease, Arch. Biochem. Biophy. 2721, 103–113.
53. Tanford, C. (1970) Protein denaturation, Adv. Protein. Chem. 24, 1–95.
54. Schellman, J. A. (1978) Solvent denaturation, Biopolymers 17, 1305–1322.
55. Pace, C. N. (1986)Determination and analysis of urea and guanidine hydrochloride denaturation curves, Meth. Enzymol. 131, 266–280.
56. Shortle, D, Meeker, A K & Freire, E. (1988) Stability mutants of staphylococcal nuclease: large compensating enthalpy-entropy changes for the reversible denaturation reaction, Biochemistry 27, 4761–4768.
57. Myers, J. K., Pace, C. N. & Scholtz, J. M. (1995) Denaturant m-values and heat capacity changes: Relation of changes in solvent accessible surface areas of protein folding, Protein Sci. 4, 2138–2148.
58. Carra, J. H., Anderson, E. A. & Privalov, P. L. (1994) Three-state thermodynamic analysis of the denaturation of staphylococcal nuclease mutants, Biochemistry 33, 10842–10850.
59. Richards, F M (1974) The interpretation of protein structures: Total volume, group volume and packing densities, J. Mol. Biol. 82, 1–14.
60. Eriksson, A E, Baase, W A, Zhang, X.J, Heinz, D W, Blaber, M, Baldwin, E & Matthews, B W (1992) The response of proteins structure to cavity creating mutations and its relationship to the hydrophobic effect, Science 255, 178–183.
61. Wynn, R, Harkins, P C, Richards, F M & Fox, R O (1997) Mobile unnatural amino acid side chains in the core of staphylococcal nuclease, Prot. Sci. 6, 1621–1626.
62. Goosens, K., Smeller, L., Frank, J. & Heremans, K. (1996) Pressure-tuning the conformation of bovine pancreatic trypsin inhibitor studied by Fourier transform infrared spectroscopy, Eur. J. Biochem., 236, 254–26
63. Peng, X., Jonas, J. & Silva, J. L. (1994) High pressure NMR study of the dissociation of arc repressor, Biochemistry, 33, 8323–832
64. Silva, J. L., Silveira, C. F., Correia Junior, A. & Pontes, L. (1992) Dissociation of a native dimer of a molten globule monomer: Effects of pressure and dilution on the dissociation equilibrium of arc repressor, J. Mol. Biol., 223, 545–55
65. Panick G Malessa R Winter R Rapp G Frye K J & Royer C A (1998) Structural characterization of the pressure denatured state and unfolding/refolding kinetics of staphylococcal nuclease by synchrotron small angle X-ray diffraction and Fourier Transform infrared spectroscopy J. Mol. Biol. 389–402
66. Flanagan, J. M., Kataoka, M., Shortle, D. & Engelman, D. M. (1992) Truncated staphylococcal nuclease in compact but disordered, Proc. Natl. Acad. Sci., 89, 748–75
67. Kataoka, M., Flanagan, J. M., Tokunaga, F. & Engelman, D. M. (1994) Use of X-ray solution scattering for a protein folding study, In: Synchrotron Radiation in the Biosciences, pp. 187–194, Chance, B., Deisenhofer, J., Ebashi, S., Goodhead, D. T., Helliwell, J. R., Huxley, H. E., Iizuka, T., Kirz, J., Mitsui, T., Rubenstein, E., Sakabe, N., Sasaki, T., Schmahl, G., Stuhrmann, H. B., Wüthrich, K. & Zaccai, G. (Eds.), Calendon Press, Oxford.
68. Desai, G., Panick, G., Winter, R. & Royer, C. A. (1998) Pressure denaturation of trp repressor, (submitted for publication).
69. Lumry, R. & Biltonen, R. (1969) in Structure and Stability of Biological Macromolecules, G. D. Fasman and S. N. Timasheff, Eds. (Marcel, Dekker, Inc., New York), vol. 2,chap. 2.
70. Gladstone, S., Laidler, K. J. & Eyring, H. (1941) in The Theory of Rate Processes, (McGraw-Hill Book Co., New York.
71. Eigen, M. & de Maeyer, L. in Techniques in Organic Chemistry, (1963) A. Weissberger, Ed., (Wiley, New York), pp. 895–1054.
72. Scalley, M. L. & Baker, D. (1997) Protein folding kinetics exhibit an Arrhenius temperature behavior when corrected for the temperature dependence of protein stability, Proc Natl. Acad. Sci. USA. 94, 10636–10640.
73. Panick, G., Vidugiris, G. J. A., Winter, R. & Royer, C. A. (1998) Exploring the temperature-pressure phase diagram of staphyiococcal nuclease (submitted for publication).
74. Nymeyer, H. Frye, K. J.. Rover, C. A., Garcia, A. E. Hummer. G. & Onuchic, J. N. Pressure probes the protein folding landscape (manuscript in preparation).
75. Kraulis. P J (1991) Molscript: A program to produce detailed and schematic plots of protein structure. J. appl. Cryst. 24, 946–950.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Royer, C.A. (1999). Pressure Denaturation of Proteins. In: Winter, R., Jonas, J. (eds) High Pressure Molecular Science. NATO Science Series, vol 358. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-4669-2_24
Download citation
DOI: https://doi.org/10.1007/978-94-011-4669-2_24
Publisher Name: Springer, Dordrecht
Print ISBN: 978-0-7923-5807-7
Online ISBN: 978-94-011-4669-2
eBook Packages: Springer Book Archive