Journal of Biomolecular NMR

, Volume 13, Issue 4, pp 381–385 | Cite as

A novel NMR experiment for the sequential assignment of proline residues and proline stretches in 13C/15N-labeled proteins

  • M.J. Bottomley
  • M.J. Macias
  • Z. Liu
  • M. Sattler


A new pulse sequence is described for the sequential assignment of proline residues in 13C/15N-labeled proteins by correlating Cδ and Cα chemical shifts of proline residues with the Hα chemical shift of the preceding residue. Notably, the experiment can provide the sequential connectivities in poly-proline stretches, which cannot be determined using standard triple resonance experiments. Excellent solvent suppression is achieved by coherence selection via a heteronuclear gradient echo. The new pulse sequence has been successfully applied to the 11 kDa HRDC domain.

proline proline-rich selective pulses sequential assignment triple resonance 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adzhubei, A.A. and Sternberg, M.J. (1993) J. Mol. Biol., 229, 472–493.Google Scholar
  2. Alexandropoulos, K., Cheng, G. and Baltimore, D. (1995) Proc. Natl. Acad. Sci. USA, 92, 3110–3114.Google Scholar
  3. Andreotti, A.H., Bunnell, S.C., Feng, S., Berg, L.J. and Schreiber, S.L. (1997) Nature, 385, 93–97.Google Scholar
  4. Bartels, C., Xia, T.-H., Billeter, M., Güntert, P., and Wüthrich, K. (1995) J. Biomol. NMR, 6, 1–10.Google Scholar
  5. Bedford, M.T., Reed, R. and Leder, P. (1998) Proc. Natl. Acad. Sci. USA, 95, 10602–10607.Google Scholar
  6. Cavanagh, J., Palmer III, A.G., Wright, P.E. and Rance, M. (1991) J. Magn. Reson., 91, 429–436.Google Scholar
  7. Chan, D.C., Bedford, M.T. and Leder, P. (1996) EMBO J., 15, 1045–1054.Google Scholar
  8. Chen, H.I., Einbond, A., Kwak, S.J., Linn, H., Koepf, E., Peterson, S., Kelly, J.W. and Sudol, M. (1997) J. Biol. Chem., 272, 17070–17077.Google Scholar
  9. Clore, G.M. and Gronenborn, A.M. (1998) Tibtech, 16, 22–34.Google Scholar
  10. Delaglio, F., Grzesiek, S., Vuister, G., Zhu, G., Pfeifer, J. and Bax, A. (1995) J. Biomol. NMR, 6, 277–293.Google Scholar
  11. Emsley, L. and Bodenhausen, G. (1990) Chem. Phys. Lett., 165, 469–476.Google Scholar
  12. Feng, S., Chen, J.K., Yu, H., Simon, J.A. and Schreiber, S.L. (1994) Science, 266, 1241–1247.Google Scholar
  13. Geen, H. and Freeman, R. (1991) J. Magn. Reson., 93, 93–141.Google Scholar
  14. Grzesiek, S. and Bax, A. (1995) J. Biomol. NMR, 6, 335–339.Google Scholar
  15. Grzesiek, S., Kuboniwa, H., Hinck, A.P. and Bax, A. (1995) J. Am. Chem. Soc., 117, 5312–5315.Google Scholar
  16. Kay, L.E., Keifer, P. and Saarinen, T. (1992) J. Am. Chem. Soc., 114, 10663–10665.Google Scholar
  17. Macias, M.J., Hyvonen, M., Baraldi, E., Schultz, J., Sudol, M., Saraste, M. and Oschkinat, H. (1996) Nature, 382, 646–649.Google Scholar
  18. Matsuo, H., Kupce, E., Li, H. and Wagner, G. (1996) J. Magn. Reson., B113, 91–96.Google Scholar
  19. McCoy, M. (1995) J. Magn. Reson., B107, 270–273.Google Scholar
  20. Morozov, V., Mushegian, A.R., Koonin, E.V. and Bork, P. (1997) Trends Biochem. Sci., 22, 417–418.Google Scholar
  21. Olejniczak, E.T. and Fesik, S.W. (1994) J. Am. Chem. Soc., 116, 2215–2216.Google Scholar
  22. Sattler, M., Schleucher, J. and Griesinger, C. (1999) Prog. NMR Spectrosc., in press.Google Scholar
  23. Schleucher, J., Sattler, M. and Griesinger, C. (1993) Angew. Chem. Int. Ed. Engl., 32, 1489–1491.Google Scholar
  24. Schleucher, J., Schwendinger, M.G., Sattler, M., Schmidt, P., Glaser, S.J., Sørensen, O.W. and Griesinger, C. (1994) J. Biomol. NMR, 4, 301–306.Google Scholar
  25. Vanhoof, G., Goossens, F., De Meester, I., Hendriks, D. and Scharpe, S. (1995) FASEB J., 9, 736–744.Google Scholar
  26. Wang, A.C., Grzesiek, S., Tschudin, R., Lodi, P.J. and Bax, A. (1995) J. Biomol. NMR, 5, 376–382.Google Scholar
  27. Williamson, M.P. (1994) Biochem. J., 297, 249–260.Google Scholar
  28. Yu, H., Chen, J.K., Feng, S., Dalgarno, D.C., Brauer, A.W. and Schreiber, S.L. (1994) Cell, 76, 933–945.Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • M.J. Bottomley
    • 1
  • M.J. Macias
    • 1
  • Z. Liu
    • 1
  • M. Sattler
    • 1
  1. 1.European Molecular Biology LaboratoryHeidelbergGermany

Personalised recommendations