Skip to main content
SpringerLink
Log in

Effect of Ficoll 70 on thermal stability and structure of creatine kinase

  • Published:
Biochemistry (Moscow) Aims and scope Submit manuscript

Abstract

The effect of Ficoll 70 on the thermal stability and structure of creatine kinase (CK) was studied using far-UV CD spectra and intrinsic fluorescence spectra. The thermal transition curves monitored by CD spectra were fitted to a two-state model using a modified form of the van’t Hoff equation to obtain the transition temperature (T m) and enthalpy change (ΔH u) of thermally induced denaturation of CK in the absence and presence of Ficoll 70. An increase in T m with constant ΔH u was observed with increasing Ficoll 70 concentration, suggesting that Ficoll 70 enhances the thermal stability of CK. Fluorescence spectral measurements confirmed this protective effect of Ficoll 70 on CK structure. In addition, we observed a crowding-induced compaction effect on the structure of both native state and thermally denatured state of CK in the presence of Ficoll 70, which is more obvious on the structure of the denatured ensemble compared to that of the native ensemble. Our observations qualitatively accord with the predictions of previously proposed crowding theory for the effect of intermolecular excluded volume on protein stability and structure. These findings imply that the effects of macromolecular crowding are essential to our understanding of protein folding and unfolding occurring in vivo.

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

Abbreviations

CK:

creatine kinase

References

  1. Anfinsen, C. B. (1973) Science, 181, 223–230.

    Article  CAS  PubMed  Google Scholar 

  2. Fulton, A. B. (1982) Cell, 30, 345–347.

    Article  CAS  PubMed  Google Scholar 

  3. Zimmerman, S. B., and Trach, S. O. (1991) J. Mol. Biol., 222, 599–620.

    Article  CAS  PubMed  Google Scholar 

  4. Ellis, R. J., and Minton, A. P. (2003) Nature, 425, 27–28.

    Article  CAS  PubMed  Google Scholar 

  5. Medalia, O., Weber, I., Frangakis, A. S., Nicastro, D., Gerisch, G., and Baumeister, W. (2002) Science, 298, 1209–1213.

    Article  CAS  PubMed  Google Scholar 

  6. Chebotareva, N. A., Kurganov, B. I., and Livanova, N. B. (2004) Biochemistry (Moscow), 69, 1239–1251.

    Article  CAS  Google Scholar 

  7. Van den Berg, B., Ellis, R. J., and Dobson, C. M. (1999) EMBO J., 18, 6927–6933.

    Article  PubMed  Google Scholar 

  8. Van den Berg, B., Wain, R., Dobson, C. M., and Ellis, R. J. (2000) EMBO J., 19, 3870–3875.

    Article  PubMed  Google Scholar 

  9. Uversky, V. N., Coopera, E. M., Bower, K. S., Li, J., and Fink, A. L. (2002) FEBS Lett., 515, 99–103.

    Article  CAS  PubMed  Google Scholar 

  10. Cheung, M. S., Klimov, D., and Thirumalai, D. (2005) Proc. Natl. Acad. Sci. USA, 102, 4753–4758.

    Article  CAS  PubMed  Google Scholar 

  11. Ai, X., Zhou, Z., Bai, Y., and Choy, W. Y. (2006) J. Am. Chem. Soc., 128, 3916–3917.

    Article  CAS  PubMed  Google Scholar 

  12. Zimmerman, S. B., and Minton, A. P. (1993) Anne. Rev. Biophys. Biomol. Struct., 22, 27–65.

    Article  CAS  Google Scholar 

  13. Minton, A. P. (2000) Curr. Opin. Struct. Biol., 10, 34–39.

    Article  CAS  PubMed  Google Scholar 

  14. Minton, A. P. (2001) J. Biol. Chem., 276, 10577–10580.

    Article  CAS  PubMed  Google Scholar 

  15. Minton, A. P. (2000) Biophys. J., 78, 101–109.

    Article  CAS  PubMed  Google Scholar 

  16. Tellam, R. L., Sculley, M. J., Nichol, L. W., and Wills, P. R. (1983) Biochem J., 213, 651–659.

    CAS  PubMed  Google Scholar 

  17. Sasahara, K., McPhie, P., and Minton, A. P. (2003) J. Mol. Biol., 326, 1227–1237.

    Article  CAS  PubMed  Google Scholar 

  18. Stagg, L., Zhang, S. Q., Cheung, M. S., and Wittung-Stafshede, P. (2007) Proc. Natl. Acad. Sci. USA, 104, 18976–18981.

    Article  CAS  PubMed  Google Scholar 

  19. Watts, D. C. (1973) in The Enzymes (Boyer, P. D., ed.) 3rd Edn., Vol. 8, Academic Press, New York, pp. 383–455.

    Google Scholar 

  20. He, H. W., Zhang, J., Zhou, H. M., and Yan, Y. B. (2005) Biophys. J., 89, 2650–2658.

    Article  CAS  PubMed  Google Scholar 

  21. Zhou, H. M., and Tsou, C. L. (1986) Biochim. Biophys. Acta, 869, 69–74.

    CAS  PubMed  Google Scholar 

  22. Zhou, H. M., Zhang, X. H., Yin, Y., and Tsou, C. L. (1993) Biochem. J., 291, 103–107.

    CAS  PubMed  Google Scholar 

  23. Yang, Y., and Zhou, H. M. (1998) Biochim. Biophys. Acta — Protein Struct. Mol. Enzymol., 1388, 190–198.

    Article  CAS  Google Scholar 

  24. Yao, Q. Z., Zhou, H. M., Hou, L. X., and Tsou, C. L. (1982) Sci. Sin. (Ser. B), 25, 1296–1302.

    CAS  Google Scholar 

  25. Yao, Q. Z., Tian, M., and Tsou, C. L. (1984) Biochemistry, 23, 2740–2744.

    Article  CAS  PubMed  Google Scholar 

  26. Zhou, H. X., and Dill, A. (2001) Biochemistry, 40, 11289–11293.

    Article  CAS  PubMed  Google Scholar 

  27. Eftink, M. R., and Ramsay, G. D. (1994) Meth. Enzymol., 240, 615–645.

    Article  PubMed  Google Scholar 

  28. Becktel, W. J., and Schellman, J. A. (1987) Biopolymers, 26, 1859–1877.

    Article  CAS  PubMed  Google Scholar 

  29. Hall, D., and Minton, A. P. (2003) Biochim. Biophys. Acta, 107, 299–316.

    Google Scholar 

  30. Ellis, R. J. (2001) Trends Biochem. Sci., 26, 597–604.

    Article  CAS  PubMed  Google Scholar 

  31. Hall, D., and Dobson, C. M. (2006) FEBS Lett., 580, 2584–2590.

    Article  CAS  PubMed  Google Scholar 

  32. Spencer, D. S., Xu, K., Logan, T. M., and Zhou, H. X. (2005) J. Mol. Biol., 351, 219–232.

    Article  CAS  PubMed  Google Scholar 

  33. Eggers, D. K., and Valentine, J. S. (2001) Protein Sci., 10, 250–261.

    Article  CAS  PubMed  Google Scholar 

  34. Karantzeni, I., Ruiz, C., Liu, C. C., and Licata, V. J. (2003) Biochem. J., 374, 785–792.

    Article  CAS  PubMed  Google Scholar 

  35. Makhatadze, G. I., and Privalov, P. L. (1995) Adv. Protein Chem., 47, 307–425.

    Article  CAS  PubMed  Google Scholar 

  36. Minton, A. P. (2005) Biophys. J., 88, 971–985.

    Article  CAS  PubMed  Google Scholar 

  37. Minton, A. P. (2005) J. Pharmacol. Sci., 94, 1668–1675.

    Article  CAS  Google Scholar 

  38. Zhou, H. X. (2004) J. Mol. Recognit., 17, 368–375.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Life Sciences, Lanzhou University, Lanzhou, 730000, Gansu, P. R. China

    Yejing Wang

  2. Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, P. R. China

    Huawei He

  3. Department of Biochemistry and Molecular Biology, Beijing Normal University, Beijing Key Laboratory, Beijing, 100875, P. R. China

    Sen Li

Authors
  1. Yejing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Huawei He
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sen Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sen Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, Y., He, H. & Li, S. Effect of Ficoll 70 on thermal stability and structure of creatine kinase. Biochemistry Moscow 75, 648–654 (2010). https://doi.org/10.1134/S0006297910050160

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0006297910050160

Key words

Search

Navigation