Differential Scanning Calorimetry of Proteins

  • Jose M. Sanchez-Ruiz
Chapter
Part of the Subcellular Biochemistry book series (SCBI, volume 24)

Abstract

Differential scanning calorimetry (DSC) is a powerful technique to characterize temperature-induced conformational changes in proteins and other biological macromolecules. In fact, DSC studies on protein thermal denaturation have played a central role in the development of current views about the factors that determine protein stability. Reviews on the various aspects of this technique are available in the literature (see, for instance, Privalov, 1979, 1982, 1989; Mateo, 1984; Sturtevant, 1987; Sanchez-Ruiz and Mateo, 1987; Freire et al., 1990) and, consequently, this chapter will mainly focus on recent developments in the field. The relevant features of the DSC experiment and the preliminary data analysis will be outlined in this section, and the well-known equilibrium thermodynamics procedures for the analysis of the DSC profiles will be briefly summarized in Section 2.

Keywords

Differential Scanning Calorimetry Heat Capacity Denature State Differential Scanning Calorimetry Experiment Excess Heat Capacity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arriaga, P., Menendez, M., Villacorta, J. M., and Laynez, J., 1992, Differential scanning calorimetric study of the thermal unfolding of β-lactamase I from Bacillus cereus. Biochemistry 31:6603–6607.PubMedGoogle Scholar
  2. Azuaga, A. I., Galisteo, M. L., Mayorga, O. L., Cortijo, M., and Mateo, P. L., 1992, Heat and cold denaturation of β-Iactoglobulin B, FEBS Lett. 309:258–260.PubMedGoogle Scholar
  3. Baldwin, R. L., 1986, Temperature dependence of the hydrophobic interaction in protein folding, Proc. Natl. Acad. Sci. USA 83:8069–8072.PubMedGoogle Scholar
  4. Baldwin, R. L., and Muller, N., 1992, Relation between the convergence temperatures T*h, and T*s in protein unfolding, Proc. Natl. Acad. Sci. USA 89:7110–7113.PubMedGoogle Scholar
  5. Becktel, W. J., and Schellman, J. A., 1987, Protein stability curves, Biopolymers 26:1859–1877.PubMedGoogle Scholar
  6. Bolen, D. W., and Santoro, M. M., 1988, Unfolding free energy determined by the linear extrapolation method. 2. Incorporation of ΔG°N-U values in a thermodynamic cycle, Biochemistry 27:8069–8074.PubMedGoogle Scholar
  7. Brandts, J. F., and Lin, L-N., 1990, Study of strong to ultratight protein interactions using differential scanning calorimetry, Biochemistry 29:6927–6940.PubMedGoogle Scholar
  8. Brandts, J. F., Hu, C. Q., and Lin, L-N., 1989, A simple model for proteins with interacting domains. Applications to scanning calorimetry data, Biochemistry 28:8588–8596.PubMedGoogle Scholar
  9. Chen, B., and Schellman, J. A., 1989, Low-temperature unfolding of a mutant of phage T4 lysozyme. 1. Equilibrium studies, Biochemistry 28:685–691.PubMedGoogle Scholar
  10. Chen, B., Baase, W. A., and Schellman, J. A., 1989, Low temperature unfolding of a mutant of phage T4 lysozyme. 2. Kinetic investigations. Biochemistry 28:691–699.PubMedGoogle Scholar
  11. Chen, B., Baase, W. A., Nicholson, H., and Schellman, J. A., 1992, Folding kinetics of T4 lysozyme and nine mutants at 12°C, Biochemistry 31:1464–1476.PubMedGoogle Scholar
  12. Chothia, C., 1976, The nature of the accessible and buried surfaces in proteins, J. Mol. Biol. 105:1–14.PubMedGoogle Scholar
  13. Conejero-Lara, F., Mateo, P. L., Aviles, F. X., and Sanchez-Ruiz, J. M. 1991a, Effect of Zn2+ on the thermal denaturation of carboxypeptidase B, Biochemistry 30:2067–2072.PubMedGoogle Scholar
  14. Conejero-Lara, F., Sanchez-Ruiz, J. M., Mateo, P. L., Burgos, F. J., Vendrell, J., and Aviles, F. X., 1991b, Differential scanning calorimetric study of carboxypeptidase B, procarboxypeptidase B and its globular activation domain, Eur. J. Biochem. 220:663–670.Google Scholar
  15. Creighton, T. E., 1990, Protein folding, Biochem. J. 270:1–16.PubMedGoogle Scholar
  16. Dill, K. A., 1985, Theory for the folding and stability of globular proteins, Biochemistry 24:1501–1509.PubMedGoogle Scholar
  17. Dill, K. A., 1990, Dominant forces in protein folding. Biochemistry 29:7133–7155.PubMedGoogle Scholar
  18. Dill, K. A., and Shortle, D., 1991, Denatured states of proteins, Annu. Rev. Biochem. 60:795–825.PubMedGoogle Scholar
  19. Doig, A. J., and Williams, D. H., 1992, Why water-soluble, compact, globular proteins have similar specific enthalpies of unfolding at 110°C, Biochemistry 31:9371–9375.PubMedGoogle Scholar
  20. Filimonov, V. V., Matveyev, S. V., Potekhin, S. A., and Privalov, P. L., 1982, Thermodynamic analysis of scanning microcalorimetric data, Mol. Biol. 16:551–562.Google Scholar
  21. Fischer, G., and Schmid, F. X., 1990, The mechanism of protein folding. Implications of in vitro refolding models for de novo protein folding and translocation in the cell. Biochemistry 29:2205–2212.PubMedGoogle Scholar
  22. Freire, E., and Biltonen, R. L., 1978, Statistical mechanical deconvolution of thermal transitions in macromolecules. I. Theory and applications to homogeneous systems, Biopolymers 17:463–479.Google Scholar
  23. Freire, E., and Murphy, K. P., 1991, The molecular basis of co-operativity in protein folding, J. Mol. Biol. 222:687–698.PubMedGoogle Scholar
  24. Freire, E., van Osdol, W. W., Mayorga, O. L., and Sanchez-Ruiz, J. ML, 1990, Calorimetrically determined dynamics of complex unfolding transitions in proteins, Annu. Rev. Biophys. Biophys. Chem. 19:159–188.PubMedGoogle Scholar
  25. Freire, E., Murphy, K. P., Sanchez-Ruiz, J. M., Galisteo, M. L., and Privalov, P. L., 1992, The molecular basis of co-operativity in protein folding. Thermodynamic dissection of interdomain interactions in phosphoglycerate kinase, Biochemistry 31:250–256.PubMedGoogle Scholar
  26. Fu, L., and Freire, E., 1992, On the origin of the enthalpy and entropy convergence temperatures in protein folding. Proc. Natl. Acad. Sci. USA 89:9335–9338.PubMedGoogle Scholar
  27. Galisteo, M. L., and Sanchez-Ruiz, J. M., 1993, Kinetic study into the irreversible thermal denaturation of bacteriorhodopsin, Eur. Biophys. J. 22:25–30.Google Scholar
  28. Galisteo, M. L., Mateo, P. L., and Sanchez-Ruiz, J. M., 1991, Kinetic study on the irreversible thermal denaturation of yeast phosphoglycerate kinase, Biochemistry 30:2061–2066.PubMedGoogle Scholar
  29. Gill, S. J., Richey, B., Bishop, G., and Wyman, J., 1985, Generalized binding phenomena in an allosteric macromolecule, Biophys. Chem. 21:1–14.PubMedGoogle Scholar
  30. Goins, B., and Freire, E., 1988, Thermal stability and intersubunit interactions of cholera toxin in solution and in association with its cell-surface receptor ganglioside G MI, Biochemistry 27:2046–2052.PubMedGoogle Scholar
  31. Green, S. M., Meeker, A. K., and Shortle, D., 1992, Contributions of the polar, uncharged amino acids to the stability of staphylococcal nuclease: Evidence for mutational effects on the free energy of the denatured state, Biochemistry 31:5717–5728.PubMedGoogle Scholar
  32. Griko, Yu. V., and Privalov, P. L., 1992, Calorimetric study of the heat and cold denaturation of β-lactoglobulin. Biochemistry 31:8810–8815.PubMedGoogle Scholar
  33. Griko, Yu. V., Privalov, P. L., Sturtevant, J. M., and Venyamov, S. Yu., 1988a. Cold denaturation of staphylococcal nuclease, Proc. Natl. Acad. Sci. USA 85:3343–3347.PubMedGoogle Scholar
  34. Griko, Yu. V., Privalov, P. L., Venyamov, S. Yu., and Kutishenko, V. P., 1988b, Thermodynamic study of the apomyoglobin structure, J. Mol. Biol. 202:128–138.Google Scholar
  35. Griko, Yu. V., Venyamov, S. Yu., and Privalov, P. L., 1989, Heat and cold denaturation of phosphoglycerate kinase (interaction of domains), FEBS Lett. 244:276–278.PubMedGoogle Scholar
  36. Guzman-Casado, M., Parody-Morreale, A., Mateo, P. L., and Sanchez-Ruiz, J. M., 1990, Differential scanning calorimetry of lobster hemocyanin, Eur. J. Biochem. 188:181–185.PubMedGoogle Scholar
  37. Hernandez-Arana, A., Rojo-Dominguez, A., Altamirano, M. M., and Calcagno, M. L., 1993, Differential scanning calorimetry of the irreversible denaturation of Escherichia coli glucosamine-6-phosphate deaminase, Biochemistry 32:3644–3648.PubMedGoogle Scholar
  38. Honig, B., Sharp, K., and Yang, A-S., 1993, Macroscopic models of aqueous solutions: Biological and chemical applications, J. Phys. Chem. 97:1101–1109.Google Scholar
  39. Hu, C-Q., Sturtevant, J. M., Thomson, J. A., Erickson, R. E., and Pace, C. N., 1992, Thermodynamics of ribonuclease T1 denaturation. Biochemistry 31:4876–4882.PubMedGoogle Scholar
  40. Jencks, W. P., 1987, Catalysis in Chemistry and Enzymology, Dover Publications Inc., New York.Google Scholar
  41. Klibanov, A. M., and Ahern, T. J., 1987, Thermal stability of proteins, in Protein Engineering (D. L. Oxender and C. F. Fox, eds.), pp. 213–218, Alan R. Liss, New York.Google Scholar
  42. Krishnan, K. S., and Brandts, J. F., 1978, Scanning calorimetry. Methods Enzymol. 49:3–14.Google Scholar
  43. Lee, B., 1991, Isoenthalpic and isoentropic temperatures and the thermodynamics of protein denaturaiton, Proc. Natl. Acad. Sci. USA 88:5154–5158.PubMedGoogle Scholar
  44. Lee, B., and Richards, F. M., 1971, The interpretation of protein structures: estimation of static accessibility, J. Mol. Biol. 55:379–400.PubMedGoogle Scholar
  45. Lepock, J. R., Rodhal, A. ML, Zhang, M. L., Heynen, M. L., Waters, B., and Cheng, K. H., 1990, Thermal denaturation of the Ca2+-ATPase of sarcoplasmic reticulum reveals two thermodynamically independent domains, Biochemistry 29:681–689.PubMedGoogle Scholar
  46. Lepock, J. R., Ritchie, K. P., Kolios, M. C, Rodahl, A. M., Heinz, K. A., and Kruuv, J., 1992, Influence of transition rates and scan rate on kinetic simulations of differential scanning calorimetry profiles of reversible and irreversible protein denaturation, Biochemistry 31:12706–12712.PubMedGoogle Scholar
  47. Lopez-Mayorga, O., and Freire, E., 1987, Dynamic analysis of differential scanning calorimetry data, Biophys. Chem. 87:87–96.Google Scholar
  48. Lumry, R., and Eyring, E., 1954, Conformation changes of proteins, J. Phys. Chem. 58:110–120.Google Scholar
  49. Makhatadze, G. I., and Privalov, P. L., 1990, Heat capacity of proteins. I. Partial molar heat capacity of individual amino acid residues in aqueous solution: Hydration effect, J. Mol. Biol. 213:375–384.PubMedGoogle Scholar
  50. Makhatadze, G. I., and Privalov, P. L., 1993, Contribution of hydration to protein folding thermodynamics. I. The enthalpy of hydration, J. Mol. Biol. 232:639–659.PubMedGoogle Scholar
  51. Manly, S. P., Matthews, K. S., and Sturtevant, J. M., 1985, Thermal denaturation of the core protein of the lac repressor, Biochemistry 24:3842–3846.PubMedGoogle Scholar
  52. Mateo, P. L., 1984, Differential scanning calorimetry of protein solutions, in Thermochemistry and Its Applications to Chemical and Biochemical Systems (R. da Silva, ed.), pp. 541–568, Reidel, Dordrecht.Google Scholar
  53. Matouschek, A., Kellis, J. T., Serrano, L., and Fersht, A. R., 1989, Mapping the transition state and pathway of protein folding by protein engineering, Nature 340:122–126.PubMedGoogle Scholar
  54. Merabet, E. K., Walker, M. C., Yuen, H. K., and Sikorski, J. A., 1993, Differential scanning calorimetric study of 5-enolpyruvoil shikimate-3-phosphate synthase and its complexes with shikimate-3-phosphate and glyphosate: Irreversible thermal transitions, Biochim. Biophys. Acta 1161:272–278.PubMedGoogle Scholar
  55. Morin, P. E., Diggs, D., and Freire, E., 1990, Thermal stability of membrane-reconstituted yeast cytochrome c oxidase. Biochemistry 29:781–788.PubMedGoogle Scholar
  56. Murphy, K. P., and Freire, E., 1992, Thermodynamics of structural stability and cooperative folding behavior in proteins. Adv. Prot. Chem. 43:313–361.Google Scholar
  57. Murphy, K. P., and Gill, S. J., 1990, Group additivity thermodynamics for dissolution of solid cyclic dipeptides in water, Thermochim. Acta 172:11–20.Google Scholar
  58. Murphy, K. P., and Gill, S. J., 1991, Solid model compounds and the thermodynamics of protein unfolding, J. Mol. Biol. 222:699–709.PubMedGoogle Scholar
  59. Murphy, K. P., Privalov, P. L., and Gill, S. J., 1990, Common features of protein unfolding and dissolution of hydrophobic compounds. Science 247:559–561.PubMedGoogle Scholar
  60. Murphy, K. P., Bhakuni, V., Xie, D., and Freire, E., 1992, Molecular basis of co-operativity in protein folding. III. Structural identification of cooperative folding units and folding intermediates, J. Mol. Biol. 227:293–306.PubMedGoogle Scholar
  61. Novokhatny, V. V., Kudinov, S. A., and Privalov, P. L., 1984, Domains in human plasminogen, J. Mol. Biol. 179:215–232.PubMedGoogle Scholar
  62. Pace, C. N., 1992, Contribution of the hydrophobic effect to globular protein stability. J. Mol. Biol. 226:29–35.PubMedGoogle Scholar
  63. Pace, C. N., and Laurents, D. V., 1989, A new method for determining the heat capacity change for protein folding, Biochemistry 28:2520–2525.PubMedGoogle Scholar
  64. Pantoliano, M. W., Withlow, M., Wood, J. F., Dodd, S. W., Hardman, K. D., Rollence, M. L., and Bryan, P. N., 1989, Large increases in general stability for subtilisin BPN’ through incremental changes in the free energy of unfolding, Biochemistry 28:7205–7213.PubMedGoogle Scholar
  65. Plaza del Pino, I. M., Pace, C. N., and Freire, E., 1992, Temperature and guanidine hydrochloride dependence of the structural stability of ribonuclease T, Biochemistry 31:11196–11202.PubMedGoogle Scholar
  66. Privalov, P. L., 1979, Stability of proteins. Small globular proteins, Adv. Prot. Chem. 33:167–241.Google Scholar
  67. Privalov, P. L., 1980, Scanning microcalorimeters for studying macromolecules, Pure Appl. Chem. 52:479–497.Google Scholar
  68. Privalov, P. L., 1982, Stability of proteins. Proteins which do not present a single cooperative system. Adv. Prot. Chem. 35:1–104.Google Scholar
  69. Privalov, P. L., 1989, Thermodynamic problems of protein structure, Anna. Rev. Biophys. Biophys. Chem. 18:47–69.Google Scholar
  70. Privalov, P. L., 1990, Cold denaturation of proteins, Crit. Rev. Biochem. Mol. Biol. 25:281–306.PubMedGoogle Scholar
  71. Privalov, P. L., and Gill, S. J., 1988, Stability of protein structure and hydrophobic interaction, Adv. Prot. Chem. 39:191–234.Google Scholar
  72. Privalov, P. L., and Khechinashvili, N. N., 1974, A thermodynamic approach to the problem of stabilization of globular protein structure: a calorimetric study, J. Mol. Biol. 86:665–684.PubMedGoogle Scholar
  73. Privalov, P. L., and Makhatadze, G. I., 1990, Heat capacity of proteins. II. Partial molar heat capacity of the unfolded polypeptide chain of proteins: protein unfolding effects, J. Mol. Biol. 213:385–391.PubMedGoogle Scholar
  74. Privalov, P. L., and Makhatadze, G. I., 1992, Contribution of hydration and non-covalent interactions to the heat capacity effect on protein unfolding, J. Mol. Biol. 224:715–723.PubMedGoogle Scholar
  75. Privalov, P. L., and Makhatadze, G. I., 1993, Contribution of hydration to protein folding thermodynamics. II. The entropy of Gibbs energy of hydration. J. Mol. Biol. 232:660–679.PubMedGoogle Scholar
  76. Privalov, P. L., Plotnikov, V. V., and Filimonov, V. V., 1975, Precision scanning microcalorimeter for the study of liquids, J. Chem. Thermodyn. 7:41–47.Google Scholar
  77. Privalov, P. L., Griko, Yu. V., Venyamov, S. Yu., and Kutyshenko, V. P., 1986, Cold denaturation of myoglobin, J. Mol. Biol. 190:487–498.PubMedGoogle Scholar
  78. Privalov, P. L., Tiktopulo, E. I., Venyaminov, S. Y., Griko, Y. V., Makhatadze, G. I., and Khechinashvili, N. N., 1989, Heat capacity and conformation of proteins in the denatured state, J. Mol. Biol. 205:737–750.PubMedGoogle Scholar
  79. Ramsay, G., and Freire, E., 1990, Linked thermal and solute perturbation analysis of cooperative domains interactions in proteins. Structural stability of diphteria toxin, Biochemistry 29:8677–8683.PubMedGoogle Scholar
  80. Robert, C. H., Gill, S. J., and Wyman, J., 1988, Quantitative analysis of linkage in macromolecules when one ligand is present in limited total quantity. Biochemistry 27:6829–6835.PubMedGoogle Scholar
  81. Sanchez-Ruiz, J. M., 1992, Theoretical analysis of Lumry-Eyring models in differential scanning calorimetry, Biophys. J. 61:921–935.PubMedGoogle Scholar
  82. Sanchez-Ruiz, J. M., and Mateo, P. L., 1987, Differential scanning calorimetry of membrane proteins. Cell Biol. Rev. 11:15–45.Google Scholar
  83. Sanchez-Ruiz, J. M., Lopez-Lacomba, J. L., Cortijo, M., and Mateo, P. L., 1988a, Differential scanning calorimetry of the irreversible thermal denaturation of thermolysin. Biochemistry 27:1648–1652.PubMedGoogle Scholar
  84. Sanchez-Ruiz, J. M., Lopez-Lacomba, J. L., Mateo, P. L., Vilanova, M., Serra, M. A., and Aviles, F. X., 1988b, Analysis of the thermal unfolding of porcine procarboxypeptidase A and its functional pieces by differential scanning calorimetry, Eur. J. Biochem. 176:225–230.PubMedGoogle Scholar
  85. Santoro, M. M., and Bolen, D. W., 1988, Unfolding free energy changes determined by the linear extrapolation method. 1. Unfolding of phenylmethanesulfonyl α-chymotrypsin using different denaturants. Biochemistry 27:8063–8068.PubMedGoogle Scholar
  86. Schellman, J. A., 1987, The thermodynamic stability of proteins, Annu. Rev. Biophys. Chem. 16:115–137.Google Scholar
  87. Scholtz, J. M., Marqusee, S., Baldwin, R. L., York, E. J., Stewart, J. M., Santoro, M., and Bolen, D. W., 1991, Calorimetric determination of the enthalpy change for the α-helix to coil transition of an alanine peptide in water, Proc. Natl. Acad. Sci. USA 88:2854–2858.PubMedGoogle Scholar
  88. Shirley, B. A., Stanssens, P., Hahn, U., and Pace, C. N., 1992, Contribution of hydrogen bonding to the conformational stability of ribonuclease T1, Biochemistry 31:725–732.PubMedGoogle Scholar
  89. Shortle, D., 1993, Denatured states of proteins and their roles in folding and stability, Curr. Opin. Struc. Biol. 3:66–74.Google Scholar
  90. Shortle, D., Stites, W. E., and Meeker, A. K., 1990, Contributions of large hydrophobic amino acids to the stability of staphylococal nuclease, Biochemistry 29:8033–8041.PubMedGoogle Scholar
  91. Shortle, D., Chan, H. S., and Dill, K., 1992, Modeling the effects of mutations on the denatured states of proteins, Protein Sci. 1:201–215.PubMedGoogle Scholar
  92. Shrake, A., and Rupley, J. A., 1973, Environment and exposure to solvent of protein atoms. Lysozyme and insuline, J. Mol. Biol. 79:351–372.PubMedGoogle Scholar
  93. Spolar, R. S., Livingstone, J. R., and Record, M. T., 1992, Use of liquid hydrocarbon and amide transfer data to estimate contributions to thermodynamic functions of protein folding from the removal of non-polar and polar surface from water. Biochemistry 31:3947–3955.PubMedGoogle Scholar
  94. Stearman, R. S., Frankel, A. D., Freire, E., Liu, B., and Pabo, C. O., 1988, Combining thermostable mutations increases the stability of λ repressor, Biochemistry 27:7571–7574.PubMedGoogle Scholar
  95. Sturtevant, J. M., 1977, Heat capacity and entropy changes in processes involving proteins, Proc. Natl. Acad. Sci. USA 74:2236–2240.PubMedGoogle Scholar
  96. Sturtevant, J. M., 1987, Biochemical applications of differential scanning calorimetry, Annu. Rev. Phys. Chem. 38:463–488.Google Scholar
  97. Takahashi, K., and Sturtevant, J. M., 1981, Thermal denaturation of Streptomyces subtilisin inhibitor, subtilisin BPN′, and the inhibitor-subtilisin complex. Biochemistry 20:6185–6190.PubMedGoogle Scholar
  98. Tamura, A., Kimura, K., Takahara, H., and Akasaka, K., 1991, Cold denaturation and heat denaturation of Streptomyces subtilisin inhibitor. 1. CD and DSC studies, Biochemistry 30:11307–11313.PubMedGoogle Scholar
  99. Wyman, J., and Gill, S. J., 1990, Binding and Linkage. Functional Chemistry of Biological Macro-molecules, University Science Books, Mill Valley, California.Google Scholar
  100. Xie, D., Bhakuni, V., and Freire, E., 1991, Calorimetric determination of the energetics of the molten globule intermediate in protein folding: Apo-α-lactalbumin, Biochemistry 30:10673–10678.PubMedGoogle Scholar
  101. Yang, A-S., Sharp, K. A., and Honig, B., 1992, Analysis of the heat capacity dependence of protein folding, J. Mol. Biol. 227:889–900.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Jose M. Sanchez-Ruiz
    • 1
  1. 1.Departamento de Quimica Fisica, Facultad de CienciasUniversidad de GranadaGranadaSpain

Personalised recommendations