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
. 1995 Jul;4(7):1315–1324. doi: 10.1002/pro.5560040707

Thermodynamics of the temperature-induced unfolding of globular proteins.

N N Khechinashvili 1, J Janin 1, F Rodier 1
PMCID: PMC2143168  PMID: 7670374

Abstract

The heat capacity, enthalpy, entropy, and Gibbs energy changes for the temperature-induced unfolding of 11 globular proteins of known three-dimensional structure have been obtained by microcalorimetric measurements. Their experimental values are compared to those we calculate from the change in solvent-accessible surface area between the native proteins and the extended polypeptide chain. We use proportionality coefficients for the transfer (hydration) of aliphatic, aromatic, and polar groups from gas phase to aqueous solution, we estimate vibrational effects, and we discuss the temperature dependence of each constituent of the thermodynamic functions. At 25 degrees C, stabilization of the native state of a globular protein is largely due to two favorable terms: the entropy of non-polar group hydration and the enthalpy of interactions within the protein. They compensate the unfavorable entropy change associated with these interactions (conformational entropy) and with vibrational effects. Due to the large heat capacity of nonpolar group hydration, its stabilizing contribution decreases quickly at higher temperatures, and the two unfavorable entropy terms take over, leading to temperature-induced unfolding.

Full Text

The Full Text of this article is available as a PDF (927.6 KB).

Selected References

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

  1. Bartunik H. D., Summers L. J., Bartsch H. H. Crystal structure of bovine beta-trypsin at 1.5 A resolution in a crystal form with low molecular packing density. Active site geometry, ion pairs and solvent structure. J Mol Biol. 1989 Dec 20;210(4):813–828. doi: 10.1016/0022-2836(89)90110-1. [DOI] [PubMed] [Google Scholar]
  2. Bernstein F. C., Koetzle T. F., Williams G. J., Meyer E. F., Jr, Brice M. D., Rodgers J. R., Kennard O., Shimanouchi T., Tasumi M. The Protein Data Bank: a computer-based archival file for macromolecular structures. J Mol Biol. 1977 May 25;112(3):535–542. doi: 10.1016/s0022-2836(77)80200-3. [DOI] [PubMed] [Google Scholar]
  3. Birktoft J. J., Blow D. M. Structure of crystalline -chymotrypsin. V. The atomic structure of tosyl- -chymotrypsin at 2 A resolution. J Mol Biol. 1972 Jul 21;68(2):187–240. doi: 10.1016/0022-2836(72)90210-0. [DOI] [PubMed] [Google Scholar]
  4. Brandts J. F., Hunt L. The thermodynamics of protein denaturation. 3. The denaturation of ribonuclease in water and in aqueous urea and aqueous ethanol mixtures. J Am Chem Soc. 1967 Sep 13;89(19):4826–4838. doi: 10.1021/ja00995a002. [DOI] [PubMed] [Google Scholar]
  5. Chen B. L., Baase W. A., Schellman J. A. Low-temperature unfolding of a mutant of phage T4 lysozyme. 2. Kinetic investigations. Biochemistry. 1989 Jan 24;28(2):691–699. doi: 10.1021/bi00428a042. [DOI] [PubMed] [Google Scholar]
  6. Connelly P., Ghosaini L., Hu C. Q., Kitamura S., Tanaka A., Sturtevant J. M. A differential scanning calorimetric study of the thermal unfolding of seven mutant forms of phage T4 lysozyme. Biochemistry. 1991 Feb 19;30(7):1887–1891. doi: 10.1021/bi00221a022. [DOI] [PubMed] [Google Scholar]
  7. Griko Y. V., Privalov P. L. Calorimetric study of the heat and cold denaturation of beta-lactoglobulin. Biochemistry. 1992 Sep 22;31(37):8810–8815. doi: 10.1021/bi00152a017. [DOI] [PubMed] [Google Scholar]
  8. Griko Y. V., Privalov P. L., Sturtevant J. M., Venyaminov SYu Cold denaturation of staphylococcal nuclease. Proc Natl Acad Sci U S A. 1988 May;85(10):3343–3347. doi: 10.1073/pnas.85.10.3343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hu C. Q., Kitamura S., Tanaka A., Sturtevant J. M. Differential scanning calorimetric study of the thermal unfolding of mutant forms of phage T4 lysozyme. Biochemistry. 1992 Feb 18;31(6):1643–1647. doi: 10.1021/bi00121a009. [DOI] [PubMed] [Google Scholar]
  10. Janin J., Wodak S. Conformation of amino acid side-chains in proteins. J Mol Biol. 1978 Nov 5;125(3):357–386. doi: 10.1016/0022-2836(78)90408-4. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Kamphuis I. G., Kalk K. H., Swarte M. B., Drenth J. Structure of papain refined at 1.65 A resolution. J Mol Biol. 1984 Oct 25;179(2):233–256. doi: 10.1016/0022-2836(84)90467-4. [DOI] [PubMed] [Google Scholar]
  13. Kanehisa M. I., Ikegami A. Structural changes and fluctuations of proteins. II. Analysis of the denaturation of globular proteins. Biophys Chem. 1977 Jan;6(2):131–149. doi: 10.1016/0301-4622(77)87003-8. [DOI] [PubMed] [Google Scholar]
  14. Khechinashvili N. N., Tsetlin V. I. Kalorimetricheskoe issledovanie teplovoi denaturatsii toksinov. Mol Biol (Mosk) 1984 May-Jun;18(3):786–791. [PubMed] [Google Scholar]
  15. Kitamura S., Sturtevant J. M. A scanning calorimetric study of the thermal denaturation of the lysozyme of phage T4 and the Arg 96----His mutant form thereof. Biochemistry. 1989 May 2;28(9):3788–3792. doi: 10.1021/bi00435a024. [DOI] [PubMed] [Google Scholar]
  16. Lee B., Richards F. M. The interpretation of protein structures: estimation of static accessibility. J Mol Biol. 1971 Feb 14;55(3):379–400. doi: 10.1016/0022-2836(71)90324-x. [DOI] [PubMed] [Google Scholar]
  17. Makhatadze G. I., Kim K. S., Woodward C., Privalov P. L. Thermodynamics of BPTI folding. Protein Sci. 1993 Dec;2(12):2028–2036. doi: 10.1002/pro.5560021204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Makhatadze G. I., Privalov P. L. Hydration effects in protein unfolding. Biophys Chem. 1994 Aug;51(2-3):291–309. doi: 10.1016/0301-4622(94)00050-6. [DOI] [PubMed] [Google Scholar]
  19. Miller S., Janin J., Lesk A. M., Chothia C. Interior and surface of monomeric proteins. J Mol Biol. 1987 Aug 5;196(3):641–656. doi: 10.1016/0022-2836(87)90038-6. [DOI] [PubMed] [Google Scholar]
  20. Murphy K. P., Privalov P. L., Gill S. J. Common features of protein unfolding and dissolution of hydrophobic compounds. Science. 1990 Feb 2;247(4942):559–561. doi: 10.1126/science.2300815. [DOI] [PubMed] [Google Scholar]
  21. Ooi T., Oobatake M. Effects of hydrated water on protein unfolding. J Biochem. 1988 Jan;103(1):114–120. doi: 10.1093/oxfordjournals.jbchem.a122215. [DOI] [PubMed] [Google Scholar]
  22. Ooi T., Oobatake M., Némethy G., Scheraga H. A. Accessible surface areas as a measure of the thermodynamic parameters of hydration of peptides. Proc Natl Acad Sci U S A. 1987 May;84(10):3086–3090. doi: 10.1073/pnas.84.10.3086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Privalov P. L., Griko YuV, Venyaminov SYu, Kutyshenko V. P. Cold denaturation of myoglobin. J Mol Biol. 1986 Aug 5;190(3):487–498. doi: 10.1016/0022-2836(86)90017-3. [DOI] [PubMed] [Google Scholar]
  25. Privalov P. L., Makhatadze G. I. Contribution of hydration and non-covalent interactions to the heat capacity effect on protein unfolding. J Mol Biol. 1992 Apr 5;224(3):715–723. doi: 10.1016/0022-2836(92)90555-x. [DOI] [PubMed] [Google Scholar]
  26. Privalov P. L., Makhatadze G. I. Contribution of hydration to protein folding thermodynamics. II. The entropy and Gibbs energy of hydration. J Mol Biol. 1993 Jul 20;232(2):660–679. doi: 10.1006/jmbi.1993.1417. [DOI] [PubMed] [Google Scholar]
  27. Privalov P. L., Makhatadze G. I. Heat capacity of proteins. II. Partial molar heat capacity of the unfolded polypeptide chain of proteins: protein unfolding effects. J Mol Biol. 1990 May 20;213(2):385–391. doi: 10.1016/S0022-2836(05)80198-6. [DOI] [PubMed] [Google Scholar]
  28. Privalov P. L. Stability of proteins: small globular proteins. Adv Protein Chem. 1979;33:167–241. doi: 10.1016/s0065-3233(08)60460-x. [DOI] [PubMed] [Google Scholar]
  29. SCHELLMAN J. A. The stability of hydrogen-bonded peptide structures in aqueous solution. C R Trav Lab Carlsberg Chim. 1955;29(14-15):230–259. [PubMed] [Google Scholar]
  30. Spolar R. S., Ha J. H., Record M. T., Jr Hydrophobic effect in protein folding and other noncovalent processes involving proteins. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8382–8385. doi: 10.1073/pnas.86.21.8382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Spolar R. S., Livingstone J. R., Record M. T., Jr Use of liquid hydrocarbon and amide transfer data to estimate contributions to thermodynamic functions of protein folding from the removal of nonpolar and polar surface from water. Biochemistry. 1992 Apr 28;31(16):3947–3955. doi: 10.1021/bi00131a009. [DOI] [PubMed] [Google Scholar]
  32. Sturtevant J. M. Heat capacity and entropy changes in processes involving proteins. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2236–2240. doi: 10.1073/pnas.74.6.2236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Swain A. L., Kretsinger R. H., Amma E. L. Restrained least squares refinement of native (calcium) and cadmium-substituted carp parvalbumin using X-ray crystallographic data at 1.6-A resolution. J Biol Chem. 1989 Oct 5;264(28):16620–16628. [PubMed] [Google Scholar]
  34. Takano T., Dickerson R. E. Conformation change of cytochrome c. II. Ferricytochrome c refinement at 1.8 A and comparison with the ferrocytochrome structure. J Mol Biol. 1981 Nov 25;153(1):95–115. doi: 10.1016/0022-2836(81)90529-5. [DOI] [PubMed] [Google Scholar]
  35. Tamura A., Kimura K., Takahara H., Akasaka K. Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 1. CD and DSC studies. Biochemistry. 1991 Nov 26;30(47):11307–11313. doi: 10.1021/bi00111a017. [DOI] [PubMed] [Google Scholar]
  36. Tanford C. Protein denaturation. C. Theoretical models for the mechanism of denaturation. Adv Protein Chem. 1970;24:1–95. [PubMed] [Google Scholar]
  37. Weaver L. H., Gray T. M., Grütter M. G., Anderson D. E., Wozniak J. A., Dahlquist F. W., Matthews B. W. High-resolution structure of the temperature-sensitive mutant of phage lysozyme, Arg 96----His. Biochemistry. 1989 May 2;28(9):3793–3797. doi: 10.1021/bi00435a025. [DOI] [PubMed] [Google Scholar]
  38. Wlodawer A., Deisenhofer J., Huber R. Comparison of two highly refined structures of bovine pancreatic trypsin inhibitor. J Mol Biol. 1987 Jan 5;193(1):145–156. doi: 10.1016/0022-2836(87)90633-4. [DOI] [PubMed] [Google Scholar]

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

RESOURCES