Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Upper limit of the time scale for diffusion and chain collapse in chymotrypsin inhibitor 2

Nature Structural Biology volume 6, pages 28–31 (1999)Cite this article

Abstract

The rates of folding of wild–type chymotrypsin inhibitor 2 (CI2) (t 1/2 = 12 ms) and of faster (t 1/2 = 2 ms) and slower (t 1/2 = 350 ms) folding mutants are accelerated in parallel by increasing concentrations of sucrose, despite the increases in viscosity. At a viscosity 26 times that of water, the folding rate constant of wild–type CI2 is accelerated four–fold (t 1/2 = 2.7 ms). From this, we can estimate that the diffusional chain collapse in CI2 occurs in less than 100 μs in water, and is not rate–determining in folding.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: a, Rate constants of folding for wild–type CI2 and mutants in the presence of sucrose.
Figure 2: Changes in the equilibrium stability and folding rate constants of the AG16/IA57 CI2 mutant with increasing sucrose concentration.
Figure 3: Rate constants for the association of barnase and barstar in the presence of the viscogenic agents sucrose and Povidone 1,300,000.

Similar content being viewed by others

References

  1. Huang, G.S. & Oas, T.G. Proc. Natl. Acad. Sci. USA 92, 6878–6882 (1995).

    Article  CAS  Google Scholar 

  2. Schindler, T.S., Herrier, M., Marahiel, M.A. & Schmid, F.X. Nature Struct. Biol. 2, 663–673 (1995).

    Article  CAS  Google Scholar 

  3. Kragelund, B.B., Robinson, C.V., Knudsen, J., Dobson, C.M. & Poulsen, F.M. Biochemistry 34, 7217–7224 (1995).

    Article  CAS  Google Scholar 

  4. Fersht, A.R. Proc. Natl. Acad. Sci. USA 92, 10869– 10873 (1995).

    Article  CAS  Google Scholar 

  5. Waldburger, C.D., Jonsson, T. & Sauer, R.T. Proc. Natl. Acad. Sci. USA 93, 2629–2634 (1996).

    Article  CAS  Google Scholar 

  6. Burton, R.E., Huang, G.S., Daugherty, M.A., Calderone, T.L. & Oas, T.G. Nature Struct. Biol. 4, 305–310 (1997).

    Article  CAS  Google Scholar 

  7. Viguera, A.R. & Serrano, L. Nature Struct. Biol. 4, 939–946 (1997).

    Article  CAS  Google Scholar 

  8. Sosnick, T.R., Mayne, L., Hiller, R. & Englander, S.W. Nature Struct. Biol. 1, 149–156 (1994).

    Article  CAS  Google Scholar 

  9. Shastry, M.C. & Roder, H. Nature Struct. Biol. 5, 385–392 (1998).

    Article  CAS  Google Scholar 

  10. Williams, S. et al. Biochemistry 35, 691– 697 (1996).

    Article  CAS  Google Scholar 

  11. Eaton, W.A., Muñoz, V., Thompson, P.A., Chan, C.K. & Hofrichter, J. Curr. Opin. Struct. Biol. 7, 10–14 (1997).

    Article  CAS  Google Scholar 

  12. Munson, M., Anderson, K.S. & Regan, L. Folding & Design 2, 77– 87 (1997).

    Article  CAS  Google Scholar 

  13. Hagen, S.J., Hofrichter, J., Szabo, A. & Eaton, W.A. Proc. Natl. Acad. Sci. USA 93, 1615– 11617 (1996).

    Article  Google Scholar 

  14. Thirumalai, D. Journal De Physique I 5, 1457–1467 (1995).

    Article  CAS  Google Scholar 

  15. Hurle, M.R., Michelotti, G.A., Crisanti, M.M. & Matthews, C.R. Proteins 2, 54–63 (1987).

    Article  CAS  Google Scholar 

  16. Chrunyk, B.A. & Matthews, C.R. Biochemistry 29, 2149–2154 (1990).

    Article  CAS  Google Scholar 

  17. Jacob, M., Schindler, T., Balbach, J. & Schmid, F.X. Proc. Natl. Acad. Sci. USA 94, 5622– 5627 (1997).

    Article  CAS  Google Scholar 

  18. Schindler, T. & Schmid, F.X. Biochemistry 35, 16833–16842 (1996).

    Article  CAS  Google Scholar 

  19. Ladurner, A.G. & Fersht, A.R. J. Mol. Biol. 273, 330–337 (1997).

    Article  CAS  Google Scholar 

  20. Li, A.J. & Daggett, V. Proc. Natl. Acad. Sci. USA 91, 10430–10434 (1994).

    Article  CAS  Google Scholar 

  21. Daggett, V., Li, A.J., Itzhaki, L.S., Otzen, D.E. & Fersht, A.R. J. Mol. Biol. 257, 430– 440 (1996).

    Article  CAS  Google Scholar 

  22. Itzhaki, L.S., Otzen, D.E. & Fersht, A.R. J. Mol. Biol. 254, 260– 288 (1995).

    Article  CAS  Google Scholar 

  23. Lazaridis, T. & Karplus . Science 278, 1928– 1931 (1997).

    Article  CAS  Google Scholar 

  24. Tan, Y.J. Ph.D. thesis. University of Cambridge (1996).

  25. Ladurner, A.G., Itzhaki, L.S., Daggett, V. & Fersht, A.R. Proc. Natl. Acad. Sci. USA 95, 8473– 8478 (1998).

    Article  CAS  Google Scholar 

  26. Stott, K., Blackburn, J.M., Butler, P.J.G. & Perutz, M.F. Proc. Natl. Acad. Sci. USA 92, 6509– 6513 (1995).

    Article  CAS  Google Scholar 

  27. Fersht, A.R., Matouschek, A. & Serrano, L. J. Mol. Biol. 224, 771– 782 (1992).

    Article  CAS  Google Scholar 

  28. Fersht, A.R., Itzhaki, L.S., elMasry, N.F., Matthews, J.M. & Otzen, D.E. Proc. Natl. Acad. Sci. USA 91, 10426–10429 (1994).

    Article  CAS  Google Scholar 

  29. Tanford, C. Adv. Prot. Chem. 24, 1–95 (1970).

    CAS  Google Scholar 

  30. Schreiber, G. & Fersht, A.R. Biochemistry 32, 5145–5150 (1993).

    Article  CAS  Google Scholar 

  31. Tsong, T.Y. Biochemistry 21, 1493–1498 (1982).

    Article  CAS  Google Scholar 

  32. Teschner, W., Rudolph, R. & Garel, J.R. Biochemistry 26, 2791– 2796 (1987).

    Article  CAS  Google Scholar 

  33. Kramers, H.A. Physica 7, 284–304 (1940).

    Article  CAS  Google Scholar 

  34. Jones, C.M. et al. Proc. Natl. Acad. Sci. USA 90, 11860–11864 (1993).

    Article  CAS  Google Scholar 

  35. Chan, C.K. et al. Proc. Natl. Acad. Sci. USA 94, 1779–1784 (1997).

    Article  CAS  Google Scholar 

  36. Chan, C.K., Hofrichter, J. & Eaton, W.A. Science 274, 628– 629 (1996).

    Article  CAS  Google Scholar 

  37. Thirumalai, D. & Woodson, S.A. Acc. Chem. Res. 29, 433–439 (1996).

    Article  CAS  Google Scholar 

  38. Camacho, C.J. & Thirumalai, D. Proc. Natl. Acad. Sci. USA 92, 1277–1281 (1995).

    Article  CAS  Google Scholar 

  39. Klimov, D.K. & Thirumalai, D. Physical Rev. Lett. 79, 317–320 (1997).

    Article  CAS  Google Scholar 

  40. Ladurner, A.G., Itzhaki, L.S. & Fersht, A.R. Folding & Design 2, 363–368 (1997).

    Article  CAS  Google Scholar 

  41. Jackson, S.E., Moracci, M., elMasry, N., Johnson, C.M. & Fersht, A.R. Biochemistry 32, 11259– 11269 (1993).

    Article  CAS  Google Scholar 

  42. McPhalen, C.A. & James, M.N.G. Biochemistry 26, 261–269 (1987).

    Article  CAS  Google Scholar 

  43. Ludvigsen, S., Shen, H., Kjær, M., Madsen, J.C., & Poulsen, F.M. J. Mol. Biol. 222, 621– 635 (1991).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank V. Daggett, D. Thirumalai and L. S. Itzhaki for discussions and communication of unpublished data, Y.M. Lam, A. Windle and A. Holmes for the use of the Ubbelohde viscometer. A.G.L. is supported by a pre–doctoral fellowship from the Boehringer Ingelheim Fonds (Stuttgart, Germany).

Author information

Author notes

  1. Andreas G. Ladurner & Alan R. Fersht

    Present address: Howard Hughes Medical Institute, Molecular and Cell Biology, University of California at Berkely, 401 Barker Hall, Berkely, California, 94720, USA

Authors and Affiliations

  1. Cambridge Unviersity Chemical Lobratory and Cambridge Center for Protein Engineering, Medical Research Council Center, Hills Road, Cambridge, CB2 2QH, UK

    Andreas G. Ladurner

Authors
  1. Andreas G. Ladurner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alan R. Fersht
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alan R. Fersht.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ladurner, A., Fersht, A. Upper limit of the time scale for diffusion and chain collapse in chymotrypsin inhibitor 2. Nat Struct Mol Biol 6, 28–31 (1999). https://doi.org/10.1038/4899

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/4899

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing