Advertisement

Journal of Biomolecular NMR

, Volume 9, Issue 3, pp 323–328 | Cite as

χ1 angle information from a simple two-dimensional NMR experiment that identifies trans 3JNCγ couplings in isotopically enriched proteins

  • Jin-Shan Hu
  • Ad Bax
Article

Abstract

New quantitative J correlation experiments are used for measuring all two- and three-bondcouplings between 15N and aliphatic side-chain carbons in proteins uniformly enriched in 13Cand 15N. Results show that 3JNCβ and 2JNCβ invariably are very small.Therefore, a simple and relatively sensitive two-dimensional spin-echo difference experimentcan be used to identify residues with a 3JNCγ coupling substantially larger than 1Hz, indicative of a trans arrangement between N and Cγ. This measurement thereforeprovides χ1 angle information for residues with an aliphatic Cγ carbon, andthereby also aids in making stereospecific assignments of Hβ resonances. Experimentsare demonstrated for ubiquitin and for a complex between calmodulin and a 26-residuepeptide.

χ1 angle Calmodulin Carbon-nitrogen J coupling Quantitative J correlation Stereospecific assignment Ubiquitin 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Supplementary material

10858_2004_199603_MOESM1_ESM.PDF (35 kb)
Supplementary Table (PDF 36 KB)

References

  1. Archer, S.J., Ikura, M., Torchia, D.A. and Bax, A. (1991) J. Magn. Reson., 95, 636–641.Google Scholar
  2. Bax, A., Mehlkopf, A.F. and Smidt, J. (1979) J. Magn. Reson., 35 167–169.Google Scholar
  3. Bax, A., Ikura, M., Kay, L.E., Torchia, D.A. and Tschudin, R. (1990) J. Magn. Reson., 86, 304–318.Google Scholar
  4. Clore, G.M., Bax, A. and Gronenborn, A.M. (1991) J. Biomol. NMR, 1, 13–22.CrossRefGoogle Scholar
  5. Eberstadt, M., Gemmecker, G., Mierke, D.F. and Kessler, H. (1995) Angew. Chem., Int. Ed. Engl., 34, 1671–1695.CrossRefGoogle Scholar
  6. Emsley, L. and Bodenhausen, G. (1990) Chem. Phys. Lett., 165, 469–476.CrossRefADSGoogle Scholar
  7. Griesinger, C., Sørensen, O.W. and Ernst, R.R. (1985) J. Am. Chem. Soc., 107, 6394–6396.CrossRefGoogle Scholar
  8. Grzesiek, S. and Bax, A. (1992) J. Magn. Reson., 96, 432–440.Google Scholar
  9. Grzesiek, S., Ikura, M., Clore, G.M., Gronenborn, A.M. and Bax, A. (1992) J. Magn. Reson., 96, 215–221.Google Scholar
  10. Güntert, P., Braun, W., Billeter, M. and Wüthrich, K. (1989) J. Am. Chem. Soc., 111, 3997–4004.CrossRefGoogle Scholar
  11. Hu, J.-S. and Bax, A. (1997) J. Am. Chem. Soc., in press.Google Scholar
  12. Hu, J.-S., Grzesiek, S. and Bax, A. (1997) J. Am. Chem. Soc., 119, 1803–1804.CrossRefGoogle Scholar
  13. Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663–10665.CrossRefGoogle Scholar
  14. Kraulis, P.J., Clore, G.M., Nilges, M., Jones, T.A., Pettersson, G., Knowles, J. and Gronenborn, A.M. (1989) Biochemistry, 28, 7241–7257.CrossRefGoogle Scholar
  15. Kuboniwa, H., Grzesiek, S., Delaglio, F. and Bax, A. (1994) J. Biomol. NMR, 4, 871–878.CrossRefGoogle Scholar
  16. Palmer, A.G., Cavanagh, J., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 93, 151–170.Google Scholar
  17. Peng, J.W. and Wagner, G. (1992) J. Magn. Reson., 98, 308–332.Google Scholar
  18. Silver, M.S., Joseph, R.I. and Hoult, D.I. (1984) J. Magn. Reson., 59, 347–351.Google Scholar
  19. Sørensen, O.W. (1990) J. Magn. Reson., 96, 433–438.Google Scholar
  20. Vuister, G.W. and Bax, A. (1993) J. Am. Chem. Soc., 115, 7772–7777.CrossRefGoogle Scholar
  21. Vuister, G.W., Wang, A.C. and Bax, A. (1993) J. Am. Chem. Soc., 115, 5334–5335.CrossRefGoogle Scholar
  22. Wagner, G., Braun, W., Havel, T.F., Schaumann, T., Gō, N. and Wüthrich, K. (1987) J. Mol. Biol., 196, 611–639.CrossRefGoogle Scholar
  23. Wand, A.J., Urbauer, J.L., McEvoy, R.P. and Bieber, R.J. (1996) Biochemistry, 35, 6116–6125.CrossRefGoogle Scholar
  24. Wang, A.C. and Bax, A. (1995) J. Am. Chem. Soc., 117, 1810–1813.CrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Jin-Shan Hu
    • 1
  • Ad Bax
    • 1
  1. 1.Laboratory of Chemical PhysicsNational Institutes of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthBethesdaU.S.A

Personalised recommendations