Skip to main content
SpringerLink
Log in

Simultaneous measurement of protein one-bond and two-bond nitrogen-carbon coupling constants using an internally referenced quantitative J-correlated [15N,1H]-TROSY-HNC experiment

  • Published:
Journal of Biomolecular NMR Aims and scope Submit manuscript

Abstract

A quantitative J-correlation pulse sequence is described that allows simultaneous determination of one-bond and two-bond nitrogen-carbon coupling constants for protonated or deuterated proteins. Coupling constants are calculated from volume ratios between cross peaks and reference axial peaks observed in a single 3D spectrum. Accurate backbone 1 J NC′, 1 J NCα, and 2 J NCα coupling constants are obtained for the two [15N;13C]-labeled, medium-sized proteins flavodoxin and xylanase and for the [2H;15N;13C]-labeled, large protein DFPase. A dependence of one-bond and two-bond J NCα values on protein backbone ψ torsion angles is readily apparent, in agreement with previously found correlations. In addition, the experiment is performed on isotropic as well as aligned protein to measure associated 15N-13C residual dipolar couplings.

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

References

  • Bax, A., Griffey, R.H. and Hawkins, B.L. (1983) J. Magn. Reson., 55, 301-315.

    Google Scholar 

  • Bax, A., Kontaxis, G. and Tjandra, N. (2001) Meth Enzymol., 339, 127-174.

    Article  Google Scholar 

  • Bax, A., Vuister, G.W., Grzesiek, S., Delaglio, F., Wang, A.C., Tschudin, R. and Zhu, G. (1994) Meth. Enzymol., 239, 79-105.

    Article  Google Scholar 

  • Bendall, M.R., Pegg, D.T. and Doddrell, D.M. (1983) J. Magn. Reson., 52, 81-117.

    Google Scholar 

  • Chou, J.J., Delaglio, F. and Bax, A. (2000) J. Biomol. NMR, 18, 101-105.

    Article  Google Scholar 

  • Choy, W.-Y., Tollinger, M., Mueller, G.A. and Kay, L.E. (2001) J. Biomol. NMR, 21, 31-40.

    Article  Google Scholar 

  • Cordier, F. and Grzesiek, S. (1999) J. Am. Chem. Soc., 121, 1601-1602.

    Article  Google Scholar 

  • Cornilescu, G., Marquardt, J.L., Ottiger, M. and Bax, A. (1998) J. Am. Chem. Soc., 121, 6836-6837.

    Article  Google Scholar 

  • Cornilescu, G., Ramirez, B.E., Frank, M.K., Clore, G.M., Gronenborn, A.M. and Bax, A. (1999) J. Am. Chem. Soc., 121, 6275-6279.

    Article  Google Scholar 

  • Curley, G.P., Carr, M.C., Mayhew, S.G. and Voordouw, G. (1991) Eur. J. Biochem., 202, 1091-1100.

    Article  Google Scholar 

  • Czisch, M. and Boelens, R. (1998) J. Magn. Reson., 134, 158-160.

    Article  ADS  Google Scholar 

  • De Alba, E. and Tjandra, N. (2002) Prog. NMR Spectrosc., 40, 175-197.

    Article  Google Scholar 

  • De Alba, E., Suzuki, M. and Tjandra, N. (2001) J. Biomol. NMR, 19, 63-67.

    Article  Google Scholar 

  • Delaglio, F., Torchia, D.A. and Bax, A. (1991) J. Biomol. NMR, 1, 439-446.

    Article  Google Scholar 

  • Dingley, A.J. and Grzesiek, S. (1998) J. Am. Chem. Soc., 120, 8293-8297.

    Article  Google Scholar 

  • Dosset, P., Hus, J.-C., Marion, D. and Blackledge, M. (2001) J. Biomol. NMR, 20, 223-231.

    Article  Google Scholar 

  • Edison, A.S., Markley, J.L. and Weinhold, F. (1994a) J. Biomol. NMR, 4, 519-542.

    Article  Google Scholar 

  • Edison, A.S., Weinhold, F., Westler, W.M. and Markley, J.L. (1994b) J. Biomol. NMR, 4, 543-551.

    Article  Google Scholar 

  • Grzesiek, S. and Bax, A. (1993) J. Am. Chem. Soc., 115, 12593-12594.

    Article  Google Scholar 

  • Hartleib, J. and Rüterjans, H. (2001) Protein Exp. Purif., 21, 210-219.

    Article  Google Scholar 

  • Heikkinen, S., Permi, P. and Kilpeläinen, I. (2001) J. Magn. Reson., 148, 53-60.

    Article  ADS  Google Scholar 

  • Hu, J.-S. and Bax, A. (1997) J. Biomol. NMR, 9, 323-328.

    Article  Google Scholar 

  • Juranić, N. and Macura, S. (2001) J. Am. Chem. Soc., 123, 4099-4100.

    Article  Google Scholar 

  • Juranić, N., Ilich, K.P. and Macura, S. (1995) J. Am. Chem. Soc., 117, 405-410.

    Article  Google Scholar 

  • Juranić, N., Likić, V.A., Prendergast, F.G. and Macura, S. (1996) J. Am. Chem. Soc., 118, 7859-7860.

    Article  Google Scholar 

  • Kay, L.E., Xu, G.Y. and Yamazaki, T. (1994) J. Magn. Reson., A109, 129-133.

    Google Scholar 

  • Knauf, M.A., Löhr, F., Curley, G.P., O’Farrell, P., Mayhew, S.G., Müller, F. and Rüterjans, H. (1993) Eur. J. Biochem., 213, 167-184.

    Article  Google Scholar 

  • Konrat, R., Muhandiram, D.R., Farrow, N.A. and Kay, L.E. (1997) J. Biomol. NMR, 9, 409-422.

    Article  Google Scholar 

  • Kupče, Ē and Freeman, R. (1995) J. Magn. Reson., A115, 273-276.

    Google Scholar 

  • Kupče, Ē, Boyd, J. and Campbell, I.D. (1995) J. Magn. Reson., B106, 300-303.

    Google Scholar 

  • Löhr, F. and Rüterjans, H. (2000) J. Magn. Reson., 146, 126-131.

    Article  ADS  Google Scholar 

  • Löhr, F., Mayhew, S.G. and Rüterjans, H. (2000) J. Am. Chem. Soc., 122, 9289-9295.

    Article  Google Scholar 

  • Löhr, F., Pérez, C., Schmidt, J.M. and Rüterjans, H. (1999) Bull. Magn. Reson., 20, 9-14.

    Google Scholar 

  • Loria, J.P., Rance, M. and Palmer III, A.G. (1999) J. Magn. Reson., 141, 180-184.

    Article  ADS  Google Scholar 

  • Matsuo, H., Kupče, Ē., Li, H. and Wagner, G. (1996) J. Magn. Reson., B111, 194-198.

    Google Scholar 

  • Meissner, A., Schulte-Herbrüggen, T. and Sørensen, O.W. (1998) J. Am. Chem. Soc., 120, 7989-7990.

    Article  Google Scholar 

  • Ottiger, M., Delaglio, F. and Bax, A. (1998) J. Magn. Reson., 131, 373-378.

    Article  ADS  Google Scholar 

  • Permi, P. and Annila, A. (2000) J. Biomol. NMR, 16, 221-227.

    Article  Google Scholar 

  • Permi, P., Rosevear, P.R. and Annila, A. (2000) J. Biomol. NMR, 17, 43-54.

    Article  Google Scholar 

  • Pervushin, K., Riek, R., Wider, G. and Wüthrich, K. (1997) Proc. Natl. Acad. Sci. USA, 94, 12366-12371.

    Article  ADS  Google Scholar 

  • Pervushin, K.V., Wider, G. and Wüthrich, K. (1998) J. Biomol. NMR, 12, 345-348.

    Article  Google Scholar 

  • Rückert, M. and Otting, G. (2000) J. Am. Chem. Soc., 122, 7793-7797.

    Article  Google Scholar 

  • Sabini, E., Sulzenbacher, G., Dauter, M., Dauter, Z., Jørgensen, P.L., Schülein, M., Dupont, C., Davies, G.J. and Wilson, K.S. (1999) Chem. Biol., 6, 483-492.

    Article  Google Scholar 

  • Salzmann, M., Pervushin, K., Wider, G., Senn, H. and Wüthrich, K. (1998) Proc. Natl. Acad. Sci. USA, 95, 13585-13590.

    Article  ADS  Google Scholar 

  • Scharff, E.I., Koepke, J., Fritzsch, G., Lücke, C. and Rüterjans, H. (2001) Structure, 9, 493-502.

    Article  Google Scholar 

  • States, D.J., Haberkorn, R.A. and Ruben, D.J. (1982) J. Magn. Reson., 48, 286-292.

    Google Scholar 

  • Stonehouse, J., Shaw, G.L., Keeler, J. and Laue, E.D. (1994) J. Magn. Reson., A107, 178-184.

    Google Scholar 

  • Tjandra, N. and Bax, A. (1997) J. Am. Chem. Soc., 119, 9576-9577.

    Article  Google Scholar 

  • Tycko, R., Pines, A. and Guckenheimer, R. (1985) J. Chem. Phys., 83, 2775-2802.

    Article  ADS  Google Scholar 

  • Vuister, G.W. and Bax, A. (1993a) J. Am. Chem. Soc., 115, 7772-7777.

    Article  Google Scholar 

  • Vuister, G.W. and Bax, A. (1993b) J. Magn. Reson., B102, 228-231.

    Google Scholar 

  • Vuister, G.W., Yamazaki, T., Torchia, D.A. and Bax, A. (1993) J. Biomol. NMR, 3, 297-306.

    Article  Google Scholar 

  • Walsh, M.A. (1994) Ph.D. Thesis, National University of Ireland.

  • Walsh, M.A., McCarthy, A., O’Farrell, P.A., McArdle, P., Cuningham, P.D., Mayhew, S.G. and Higgins, T.M. (1998) Eur. J. Biochem., 258, 362-371.

    Article  Google Scholar 

  • Wang, Y.-X., Jacob, J., Cordier, F., Wingfield, P., Stahl, S.J., Lee-Huang, S., Torchia, D., Grzesiek, S. and Bax, A. (1999) J. Biomol. NMR, 14, 181-184.

    Article  MATH  Google Scholar 

  • Weigelt, J. (1998) J. Am. Chem. Soc., 120, 10778-10779.

    Article  Google Scholar 

  • Wirmer, J. and Schwalbe, H. (2002) J. Biomol. NMR, 23, 47-55.

    Article  Google Scholar 

  • Zweckstetter, M. and Bax, A. (2000) J. Am. Chem. Soc., 122, 3791-3792.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Biophysikalische Chemie, J.W. Goethe-Universität, Marie-Curie-Str. 9, D-60439, Frankfurt am Main, Germany

    Hans L.J. Wienk, Mitcheell M. Martínez, Gary N. Yalloway, Carlos Pérez, Heinz Rüterjans & Frank Löhr

  2. Research School of Biosciences, University of Kent at Canterbury, Canterbury, Kent, CT2 7NJ, U.K

    Jürgen M. Schmidt

Authors
  1. Hans L.J. Wienk
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mitcheell M. Martínez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gary N. Yalloway
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jürgen M. Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Carlos Pérez
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Heinz Rüterjans
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Frank Löhr
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Frank Löhr.

Electronic Supplementary Material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wienk, H.L., Martínez, M.M., Yalloway, G.N. et al. Simultaneous measurement of protein one-bond and two-bond nitrogen-carbon coupling constants using an internally referenced quantitative J-correlated [15N,1H]-TROSY-HNC experiment. J Biomol NMR 25, 133–145 (2003). https://doi.org/10.1023/A:1022233103990

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022233103990

Search

Navigation