Letters in Peptide Science

, Volume 4, Issue 4–6, pp 245–249 | Cite as

Conformational study of a native monodisulfide bridge analogue of EETI II

  • Annie Heitz
  • Dung Le-Nguyen
  • Bertrand Castro
  • Laurent Chiche
Article

Summary

Trypsin inhibitor EETI II, possessing six cysteines engaged in three disulfide bridges, shares a common structural motif with other proteins of different origins and functions. To understand the principles that govern folding of this largely distributed basic scaffold, mainly composed of a small triple-stranded β-sheet, we have studied different stages in the folding of EETI II. The conformational properties of a synthetic analogue of EETI II possessing only one native (15–27) disulfide bridge were investigated with the combined use of1H NMR and molecular modelling. Although two native-like reverse turns were observed, formation of β-sheet could not be evidenced in the one disulfide analogue, while the motif has been shown to be present in a folding intermediate with two native disulfide bridges (9–21 and 15–27). These results suggest that the structural motif requires stabilisation by two disulfide bridges.

Keywords

Folding Molecular modelling NMR Squash trypsin inhibitor 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Heitz, A., Chiche, L., Le-Nguyen D. and Castro, B., Biochemistry, 28 (1989) 2392.PubMedCrossRefGoogle Scholar
  2. 2.
    Chiche, L., Gaboriaud, C., Heitz, A., Mornon, J.P., Castro, B. and Kollman, P.A., Proteins, 6 (1989) 405.PubMedCrossRefGoogle Scholar
  3. 3.
    Chiche, L., Heitz, A., Padilla, A., Le-Nguyen, D. and Castro, B., Protein Eng., 6 (1993) 675.PubMedGoogle Scholar
  4. 4.
    Seater, O., Craik, D.J., Campbell, I.D., Sletten, K., Juul, J. and Norman, D.G., Biochemistry, 34 (1995) 4147.CrossRefGoogle Scholar
  5. 5.
    Pallaghy, P.K., Nielsen, K.J., Craik, D.J. and Norton, R.S., Protein Sci., 3 (1994) 1833.PubMedCrossRefGoogle Scholar
  6. 6.
    Le-Nguyen, D., Heitz, A., Chiche, L., Castro, B., Boigegrain, R.A., Favel, A. and Coletti-Previero, M.A., Biochimie, 72 (1990) 431.PubMedCrossRefGoogle Scholar
  7. 7.
    Le-Nguyen, D., Heitz, A., Chiche, L., El Hajji, M. and Castro, B., Protein Sci., 2 (1993) 165.PubMedCrossRefGoogle Scholar
  8. 8.
    Heitz, A., Chiche, L., Le-Nguyen, D. and Castro, B., Eur. J. Biochem., 233 (1995) 837.PubMedCrossRefGoogle Scholar
  9. 9.
    Le-Nguyen, D., Heitz, A. and Castro, B., J. Chem. Soc., Perkin Trans I, (1987) 1915.CrossRefGoogle Scholar
  10. 10.
    Wüthrich, K., NMR of Proteins and Nucleic Acids, Wiley, New York, NY, U.S.A., 1986.Google Scholar
  11. 11.
    Wüthrich, K., Billeter, M. and Braun, W., J. Mol. Biol., 169 (1983) 949.PubMedGoogle Scholar
  12. 12.
    Güntert, P., Mumenthaler, C. and Wüthrich, K., Abstracts XVIIth International Conference on Magnetic Resonance in Biological Systems, Keystone/Colorado, U.S.A., August 23–28, 1996.Google Scholar
  13. 13.
    Kraulis, P., J. Appl. Crystallogr., 24 (1991) 946.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Annie Heitz
    • 1
  • Dung Le-Nguyen
    • 2
  • Bertrand Castro
    • 3
  • Laurent Chiche
    • 1
  1. 1.Centre de Biochimie StructuraleCNRS-INSERM, Faculté de PharmacieMontpellier Cedex 1France
  2. 2.Unité 376 INSERMCHU Arnaud de VilleneuveMontpellier Cedex 05France
  3. 3.Sanofi-ChimieGentilly CedexFrance

Personalised recommendations