Skip to main content
NCBI home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1999 Jul;8(7):1469–1474. doi: 10.1110/ps.8.7.1469

Pressure-induced unfolding of lysozyme in aqueous guanidinium chloride solution.

K Sasahara 1, K Nitta 1
PMCID: PMC2144367  PMID: 10422835

Abstract

The pressure-induced unfolding of lysozyme was investigated in an aqueous guanidinium chloride solution by means of ultraviolet spectroscopy. Assuming a two-state transition model, volume changes were calculated from the slope of free energy vs. pressure plots over a temperature range of 10 to 60 degrees C. Between 25 and 60 degrees C, almost constant volume changes were observed in the transition region, which was reflected in almost identical slopes of the free energy change vs. pressure plots. On the other hand, the different slopes were observed in the pressure dependence of free energy change at temperatures lower than 25 degrees C. These data were interpreted as suggesting that a two-state model is not appropriate at low temperature, but instead one or more intermediates are present under these conditions. The volume changes for unfolding became less negative at temperatures higher than 25 degrees C.

Full Text

The Full Text of this article is available as a PDF (454.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aune K. C., Tanford C. Thermodynamics of the denaturation of lysozyme by guanidine hydrochloride. II. Dependence on denaturant concentration at 25 degrees. Biochemistry. 1969 Nov;8(11):4586–4590. doi: 10.1021/bi00839a053. [DOI] [PubMed] [Google Scholar]
  2. Brandts J. F., Oliveira R. J., Westort C. Thermodynamics of protein denaturation. Effect of pressu on the denaturation of ribonuclease A. Biochemistry. 1970 Feb 17;9(4):1038–1047. doi: 10.1021/bi00806a045. [DOI] [PubMed] [Google Scholar]
  3. Chalikian T. V., Bresiauer K. J. On volume changes accompanying conformational transitions of biopolymers. Biopolymers. 1996 Nov;39(5):619–626. doi: 10.1002/(sici)1097-0282(199611)39:5<619::aid-bip1>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  4. Chen L., Hodgson K. O., Doniach S. A lysozyme folding intermediate revealed by solution X-ray scattering. J Mol Biol. 1996 Sep 6;261(5):658–671. doi: 10.1006/jmbi.1996.0491. [DOI] [PubMed] [Google Scholar]
  5. Creighton T. E. Protein folding. Biochem J. 1990 Aug 15;270(1):1–16. doi: 10.1042/bj2700001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dill K. A., Shortle D. Denatured states of proteins. Annu Rev Biochem. 1991;60:795–825. doi: 10.1146/annurev.bi.60.070191.004051. [DOI] [PubMed] [Google Scholar]
  7. Franks F. Protein destabilization at low temperatures. Adv Protein Chem. 1995;46:105–139. doi: 10.1016/s0065-3233(08)60333-2. [DOI] [PubMed] [Google Scholar]
  8. Frye K. J., Royer C. A. Probing the contribution of internal cavities to the volume change of protein unfolding under pressure. Protein Sci. 1998 Oct;7(10):2217–2222. doi: 10.1002/pro.5560071020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Griko Y. V., Freire E., Privalov P. L. Energetics of the alpha-lactalbumin states: a calorimetric and statistical thermodynamic study. Biochemistry. 1994 Feb 22;33(7):1889–1899. doi: 10.1021/bi00173a036. [DOI] [PubMed] [Google Scholar]
  10. Gross M., Jaenicke R. Proteins under pressure. The influence of high hydrostatic pressure on structure, function and assembly of proteins and protein complexes. Eur J Biochem. 1994 Apr 15;221(2):617–630. doi: 10.1111/j.1432-1033.1994.tb18774.x. [DOI] [PubMed] [Google Scholar]
  11. Haezebrouck P., Joniau M., Van Dael H., Hooke S. D., Woodruff N. D., Dobson C. M. An equilibrium partially folded state of human lysozyme at low pH. J Mol Biol. 1995 Feb 24;246(3):382–387. doi: 10.1006/jmbi.1994.0093. [DOI] [PubMed] [Google Scholar]
  12. Hawley S. A. Reversible pressure--temperature denaturation of chymotrypsinogen. Biochemistry. 1971 Jun 22;10(13):2436–2442. doi: 10.1021/bi00789a002. [DOI] [PubMed] [Google Scholar]
  13. Hummer G., Garde S., García A. E., Paulaitis M. E., Pratt L. R. The pressure dependence of hydrophobic interactions is consistent with the observed pressure denaturation of proteins. Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1552–1555. doi: 10.1073/pnas.95.4.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ikeguchi M., Kuwajima K., Mitani M., Sugai S. Evidence for identity between the equilibrium unfolding intermediate and a transient folding intermediate: a comparative study of the folding reactions of alpha-lactalbumin and lysozyme. Biochemistry. 1986 Nov 4;25(22):6965–6972. doi: 10.1021/bi00370a034. [DOI] [PubMed] [Google Scholar]
  15. Janin J. Surface and inside volumes in globular proteins. Nature. 1979 Feb 8;277(5696):491–492. doi: 10.1038/277491a0. [DOI] [PubMed] [Google Scholar]
  16. KAUZMANN W. Some factors in the interpretation of protein denaturation. Adv Protein Chem. 1959;14:1–63. doi: 10.1016/s0065-3233(08)60608-7. [DOI] [PubMed] [Google Scholar]
  17. Kim P. S., Baldwin R. L. Intermediates in the folding reactions of small proteins. Annu Rev Biochem. 1990;59:631–660. doi: 10.1146/annurev.bi.59.070190.003215. [DOI] [PubMed] [Google Scholar]
  18. Kuwajima K., Hiraoka Y., Ikeguchi M., Sugai S. Comparison of the transient folding intermediates in lysozyme and alpha-lactalbumin. Biochemistry. 1985 Feb 12;24(4):874–881. doi: 10.1021/bi00325a010. [DOI] [PubMed] [Google Scholar]
  19. Lee J. C., Timasheff S. N. Partial specific volumes and interactions with solvent components of proteins in guanidine hydrochloride. Biochemistry. 1974 Jan 15;13(2):257–265. doi: 10.1021/bi00699a005. [DOI] [PubMed] [Google Scholar]
  20. Li T. M., Hook J. W., 3rd, Drickamer H. G., Weber G. Plurality of pressure-denatured forms in chymotrypsinogen and lysozyme. Biochemistry. 1976 Dec 14;15(25):5571–5580. doi: 10.1021/bi00670a023. [DOI] [PubMed] [Google Scholar]
  21. Makhatadze G. I., Privalov P. L. Protein interactions with urea and guanidinium chloride. A calorimetric study. J Mol Biol. 1992 Jul 20;226(2):491–505. doi: 10.1016/0022-2836(92)90963-k. [DOI] [PubMed] [Google Scholar]
  22. Miranker A., Radford S. E., Karplus M., Dobson C. M. Demonstration by NMR of folding domains in lysozyme. Nature. 1991 Feb 14;349(6310):633–636. doi: 10.1038/349633a0. [DOI] [PubMed] [Google Scholar]
  23. Nash D. P., Jonas J. Structure of pressure-assisted cold denatured lysozyme and comparison with lysozyme folding intermediates. Biochemistry. 1997 Nov 25;36(47):14375–14383. doi: 10.1021/bi970881v. [DOI] [PubMed] [Google Scholar]
  24. Nozaki Y. The preparation of guanidine hydrochloride. Methods Enzymol. 1972;26:43–50. doi: 10.1016/s0076-6879(72)26005-0. [DOI] [PubMed] [Google Scholar]
  25. Privalov P. L., Gill S. J. Stability of protein structure and hydrophobic interaction. Adv Protein Chem. 1988;39:191–234. doi: 10.1016/s0065-3233(08)60377-0. [DOI] [PubMed] [Google Scholar]
  26. Privalov P. L., Tiktopulo E. I., Venyaminov SYu, Griko YuV, Makhatadze G. I., Khechinashvili N. N. Heat capacity and conformation of proteins in the denatured state. J Mol Biol. 1989 Feb 20;205(4):737–750. doi: 10.1016/0022-2836(89)90318-5. [DOI] [PubMed] [Google Scholar]
  27. Radford S. E., Dobson C. M., Evans P. A. The folding of hen lysozyme involves partially structured intermediates and multiple pathways. Nature. 1992 Jul 23;358(6384):302–307. doi: 10.1038/358302a0. [DOI] [PubMed] [Google Scholar]
  28. Rose G. D., Geselowitz A. R., Lesser G. J., Lee R. H., Zehfus M. H. Hydrophobicity of amino acid residues in globular proteins. Science. 1985 Aug 30;229(4716):834–838. doi: 10.1126/science.4023714. [DOI] [PubMed] [Google Scholar]
  29. Samarasinghe S. D., Campbell D. M., Jonas A., Jonas J. High-resolution NMR study of the pressure-induced unfolding of lysozyme. Biochemistry. 1992 Sep 1;31(34):7773–7778. doi: 10.1021/bi00149a005. [DOI] [PubMed] [Google Scholar]
  30. Skerjanc J., Lapanje S. A dilatometric study of the denaturation of lysozyme by guanidine hydrochloride and hydrochloric acid. Eur J Biochem. 1972 Jan 31;25(1):49–53. doi: 10.1111/j.1432-1033.1972.tb01665.x. [DOI] [PubMed] [Google Scholar]
  31. Tamura A., Kimura K., Akasaka K. Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 2. 1H NMR studies. Biochemistry. 1991 Nov 26;30(47):11313–11320. doi: 10.1021/bi00111a018. [DOI] [PubMed] [Google Scholar]
  32. Tanford C. Protein denaturation. Adv Protein Chem. 1968;23:121–282. doi: 10.1016/s0065-3233(08)60401-5. [DOI] [PubMed] [Google Scholar]
  33. Wolfenden R., Andersson L., Cullis P. M., Southgate C. C. Affinities of amino acid side chains for solvent water. Biochemistry. 1981 Feb 17;20(4):849–855. doi: 10.1021/bi00507a030. [DOI] [PubMed] [Google Scholar]
  34. Yamaguchi T., Yamada H., Akasaka K. Thermodynamics of unfolding of ribonuclease A under high pressure. A study by proton NMR. J Mol Biol. 1995 Jul 28;250(5):689–694. doi: 10.1006/jmbi.1995.0408. [DOI] [PubMed] [Google Scholar]
  35. Zhang J., Peng X., Jonas A., Jonas J. NMR study of the cold, heat, and pressure unfolding of ribonuclease A. Biochemistry. 1995 Jul 11;34(27):8631–8641. doi: 10.1021/bi00027a012. [DOI] [PubMed] [Google Scholar]
  36. Zipp A., Kauzmann W. Pressure denaturation of metmyoglobin. Biochemistry. 1973 Oct 9;12(21):4217–4228. doi: 10.1021/bi00745a028. [DOI] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES