Skip to main content
SpringerLink
Log in

Binding isotherm determination by isothermal titration calorimetry

  • Published:
Journal of Thermal Analysis and Calorimetry Aims and scope Submit manuscript

Abstract

A simple method for determination of binding isotherm in the protein-ligand interaction was introduced using isothermal titration calorimetric data. This general method was applied to the study of the interaction of myelin basic protein (MBP) from bovine central nervous system with divalent copper ion at 27°C in Tris buffer solution at pH=7.2. The binding isotherm for copper-MBP interaction is easily obtained by carrying out titration calorimetric experiment in two different concentrations of MBP. MBP has two binding sites for copper ion, which show positive cooperativity in its sites. The intrinsic association equilibrium constants are 0.083 and 1.740 ?M-1 in the first and second binding sites, respectively. Hence, occupation of the first site has produced an appreciable enhancement 21 of the binding affinity of the second site. The molar enthalpies of binding are -13.5 and -14.8 kJ mol-1 in the first and second binding sites, respectively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. J. T. Edsall and H. Gutfreund, Biothermodynamics, Wiley, New York 1984, Chapter 6.

    Google Scholar 

  2. L. Peng, L. Yi, H. Jia, D. Fengjiao, P. Daiwen and Q. Songsheng, J. Therm. Anal. Cal., 73 (2003) 843.

    Article  CAS  Google Scholar 

  3. I. Wadso, Thermochim. Acta, 394 (2002) 305.

    Article  CAS  Google Scholar 

  4. E. Freire, O. L. Mayorga and M. Straume, Anal. Chem., 62 (1990) 950A.

    CAS  Google Scholar 

  5. S. Leavitt and E. Freire, Current Opinion in Structural Biology, 11 (2001) 560.

    Article  CAS  Google Scholar 

  6. A. A. Saboury, A. K. Bordbar and A. A. Moosavi-Movahedi, Bull. Chem. Soc. Jpn., 69 (1996) 3031.

    Article  CAS  Google Scholar 

  7. A. A. Saboury, M. U. Dahut, S. Ghobadi, J. Chamani and A. A. Moosavi-Movahedi, J. Chin. Chem. Soc., 45 (1998) 667.

    CAS  Google Scholar 

  8. A. A. Saboury, Thermochim. Acta, 320 (1998) 97.

    Article  CAS  Google Scholar 

  9. S. Z. Bathaie, A. A. Moosavi-Movahedi and A. A. Saboury, Nuc. Acid Res., 27 (1999) 1001.

    Article  CAS  Google Scholar 

  10. A. A. Saboury and F. Karbassi, Thermochim. Acta, 362 (2000) 121.

    Article  CAS  Google Scholar 

  11. A. Arabzadeh, S. Z. Bathaie, H. Farsam, M. Amanlou, A. A. Saboury and A. A. Moosavi-Movahedi, Inter. J. Pharmac., 237 (2002) 47.

    Article  CAS  Google Scholar 

  12. T. D. Spurway, C. Morland, A. Cooper, I. Sumner, G. P. Hazlewood, A. G. O'Donnell, R. W. Pickersgill and H. J. Gilbert, J. Biol. Chem., 272 (1997) 17523.

    Article  CAS  Google Scholar 

  13. C. Hammanu, A. Cooper and D. M. Lilley, Biochemistry, 40 (2001) 1423.

    Article  Google Scholar 

  14. O. A. Amire, J. Masoudy, A.A. Saboury and A.A. Moosavi-Movahedi, Thermochim. Acta, 303 (1997) 219.

    Article  CAS  Google Scholar 

  15. A. K. Bordbar, A. A. Moosavi-Movahedi and A. A. Saboury, Thermochim. Acta, 287 (1996) 343.

    Article  CAS  Google Scholar 

  16. A. A. Saboury, A. K. Bordbar and A. A. Moosavi-Movahedi, J. Chem. Thermodyn., 28 (1996) 1077.

    Article  CAS  Google Scholar 

  17. K. Nazari, A. A. Saboury and A. A. Moosavi-Movahedi, Thermochim. Acta, 302 (1997) 131.

    Article  CAS  Google Scholar 

  18. A. A. Moosavi-Movahedi and A. A. Saboury, Jour. Chem. Soc. Pak., 21 (1999) 248.

    CAS  Google Scholar 

  19. J. Chamani, A. A. Moosavi-Movahedi, A. A. Saboury, M. Gharanfoli and G. H. Hakimelahi, J. Chem. Thermodyn., 35 (2003) 199.

    Article  CAS  Google Scholar 

  20. A. A. Saboury and A. A. Moosavi-Movahedi, J. Enzyme Inhibition, 12 (1997) 273.

    CAS  Google Scholar 

  21. A. A. Saboury, Indian J. Biochem. Biophys., 37 (2000) 347.

    CAS  Google Scholar 

  22. A. A. Saboury, A. Divsalar, G. Ataie, A. A. Moosavi-Movahedi, M. R. Housaindokht and G. H. Hakimelahi, J. Biochem. Mol. Biol., 35 (2002) 302.

    CAS  Google Scholar 

  23. N. Sarri-Sarraf, A. A. Saboury and A. A. Moosavi-Movahedi, J. Enz. Inhib. Med. Chem., 17 (2002) 203.

    Article  Google Scholar 

  24. M. Angberg, C. Nystrom and S. Castensson, Acta Pharm. Suec., 25 (1988) 307.

    CAS  Google Scholar 

  25. L. D. Hansen, E. A. Lewis, D. J. Eatough, R. G. Bergstrom and D. DeGraft-Johnson, Pharm. Res., 6 (1989) 20.

    Article  CAS  Google Scholar 

  26. M. J. Koenigbaure, S. H. Brooks, G. Rullo and R. A. Couch, Pharm. Res., 9 (1992) 939.

    Article  Google Scholar 

  27. A. A. Saboury, M. Miroliae, M. Nemat-Gorgani and A. A. Moosavi-Movahedi, Thermochim. Acta, 326 (1999) 127.

    Article  CAS  Google Scholar 

  28. T. F. Bolles and R. S. Drago, J. Am. Chem. Soc., 87 (1965) 5015.

    Article  CAS  Google Scholar 

  29. M. A. Laandav, M. N. Markovich and L. A. Pyruzyan, Biochim. Biophys. Acta, 493 (1977) 1.

    Google Scholar 

  30. A. Chen and I. Wadso, Biochem. Biophys. Meth., 6 (1982) 307.

    Article  CAS  Google Scholar 

  31. T. Wiseman, S. Williston, J. F. Brandts and L. Lin, Anal. Biochem., 179 (1989) 131.

    Article  CAS  Google Scholar 

  32. A. A. Saboury, S. Saeidian, M. H. Sanati, A. A. Moosavi-Movahedi and F. Alasti, J. Chin. Chem. Soc., 48 (2001) 827.

    CAS  Google Scholar 

  33. A. A. Saboury, N. Sarri-Sarraf and S. Saeidian, Thermochim Acta, 381 (2002) 147.

    Article  CAS  Google Scholar 

  34. P. R. Connelly, R. Varadarajan, J. M. Sturtevant and F. M. Richards, Biochemistry, 29 (1990) 6108.

    Article  CAS  Google Scholar 

  35. J. E. Wilson and A. Chin, Anal. Biochem., 193 (1991) 16.

    Article  CAS  Google Scholar 

  36. F. Zhang and E. S. Rowe, Biochemistry, 31 (1992) 2005.

    Article  CAS  Google Scholar 

  37. D. Hamada, S. Kidokoro, H. Fukada, K. Takahashi and Y. Goto, Proc. Natl. Acad. Sci. USA, 91 (1994) 10325.

    Article  CAS  Google Scholar 

  38. R. Ramkumar, A. Surolia and S. K. Podder, Biochem. J., 308 (1995) 237.

    CAS  Google Scholar 

  39. C. S. Raman, M. J. Allan and B. Nall, Biochemistry, 34 (1995) 5831.

    Article  CAS  Google Scholar 

  40. M. C. Chervenak and E. J. Toone, Biochemistry, 34 (1995) 5685.

    Article  CAS  Google Scholar 

  41. A. A. Saboury, Jour. Chem. Soc. Pak., 22 (2000) 204.

    Google Scholar 

  42. A. A. Saboury, J. Therm. Anal. Cal., 72 (2003) 93.

    Article  CAS  Google Scholar 

  43. A. A. Saboury, Biologia, 57 (2002) 221.

    CAS  Google Scholar 

  44. M. Ghadermarzi, A. A. Saboury and A. A. Moosavi-Movahedi, Polish J. Chem., 72 (1998) 2024.

    CAS  Google Scholar 

  45. A. A. Saboury, J. Chem. Thermodyn., 17 (2003) 1975.

    Article  Google Scholar 

  46. C. M. Deber and S. J. Reynolds, Clin. Biochem., 24 (1991) 113.

    Article  CAS  Google Scholar 

  47. G. Scatchard, Ann. New York Acad. Sci., 50 (1949) 660.

    Google Scholar 

  48. A. A. Saboury and A. A. Moosavi-Movahedi, Biochem. Educ., 22 (1994) 48.

    Article  Google Scholar 

  49. A. K. Bordbar, A. A. Saboury and A. A. Moosavi-Movahedi, Biochem. Educ., 24 (1996) 172.

    Article  CAS  Google Scholar 

  50. A. V. Hill, J. Physiol., 40 (1910) IV.

    Google Scholar 

  51. M. L. James, G. M. Smith and J. C. Wolford, Applied Numerical Methods for Digital Computer, 3rd Ed., Harper and Row, New York 1985.

    Google Scholar 

  52. W. C. Galley, M. Bouvier, S.-D. Clas, G. R. Brown and L. E. St-Pierre, Biopolymers, 27 (1988) 79.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Biochemistry and Biophysics, University of Tehran Tehran, Iran E-mail

    A. A. Saboury

Authors
  1. A. A. Saboury
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Saboury, A.A. Binding isotherm determination by isothermal titration calorimetry. Journal of Thermal Analysis and Calorimetry 77, 997–1004 (2004). https://doi.org/10.1023/B:JTAN.0000041675.76664.07

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/B:JTAN.0000041675.76664.07

Search

Navigation