Skip to main content
SpringerLink
Log in

Heteronuclear relayed E.COSY revisited: Determination of 3J(Hα,Cγ) couplings in Asx and aromatic residues in proteins

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

Abstract

Constant-time 3D heteronuclear relayed E.COSY [Schmidt et al. (1996) J. Biomol. NMR, 7, 142–152], as based on generic 2D small-flip-angle HMQC-COSY [Schmidt et al. (1995) J. Biomol. NMR, 6, 95–105], has been modified to allow for quantitative determination of heteronuclear three-bond 3 J(Hα,Cγ) couplings. The method is applicable to amino acid spin topologies with carbons in the γ position which lack attached protons, i.e. to asparagine, aspartate, and aromatic residues in uniformly 13C-enriched proteins. The pulse sequence critically exploits heteronuclear triple-quantum coherence (HTQC) of CH2 moieties involving geminal Hβ proton pairs, taking advantage of improved multiple-quantum relaxation properties, at the same time avoiding scalar couplings between those spins involved in multiple-quantum coherence, thus yielding E.COSY-type multiplets with a splitting structure that is simpler than with the original scheme. Numerical least-squares 2D line-shape simulation is used to extract 3 J(Hα,Cγ) coupling constants which are of relevance to side-chain χ1 dihedral-angle conformations in polypeptides. Methods are demonstrated with recombinant 15N,13C-enriched ribonuclease T1 and Desulfovibrio vulgaris flavodoxin with bound oxidized FMN.

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

  • Aue, W.P., Bartholdi, E. and Ernst, R.R. (1976) J. Chem. Phys., 64 2229–2246.

    Google Scholar 

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

    Google Scholar 

  • Blümel, M., Schmidt, J.M., Löhr, F. and Rüterjans, H. (1998) Eur.Biophys. J., 27, 321–334.

    Google Scholar 

  • DeMarco, A., Llinás, M. and Wüthrich, K. (1978) Biopolymers, 17, 617–636.

    Google Scholar 

  • DeMarco, A. and Llinás, M. (1979) Biochemistry, 18, 3846–3854.

    Google Scholar 

  • Eggenberger, U., Schmidt, P., Sattler, M., Glaser, S.J. and Griesinger, C. (1992) J. Magn. Reson., 100, 604–610.

    Google Scholar 

  • Emsley, L. and Bodenhausen, G. (1990) Chem. Phys. Lett., 165, 469–476.

    Google Scholar 

  • Ernst, R.R., Bodenhausen, G. and Wokaun, A. (1987) Principles of Nuclear Magnetic Resonance in One and Two Dimensions, Clarendon Press, Oxford, U.K.

    Google Scholar 

  • Griesinger, C., Sørensen, O.W. and Ernst, R.R. (1986) J. Chem. Phys., 85, 6837–6852.

    Google Scholar 

  • Griffey, R.H. and Redfield, A.G. (1987) Quart. Rev. Biophys., 19, 51–82.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1993) J. Biomol. NMR, 3, 185–204.

    Google Scholar 

  • Grzesiek, S. and Bax, A. (1995) J. Biomol. NMR, 6, 335–339.

    Google Scholar 

  • Gschwind, R.M., Gemmecker, G. and Kessler, H. (1998) J. Biomol. NMR, 11, 191–198.

    Google Scholar 

  • Karimi-Nejad, Y. (1994) Thesis, University of Cologne, Germany.

    Google Scholar 

  • Logan, T.M., Olejniczak, E.T., Xu, R.X. and Fesik, S.W. (1992) FEBS Lett., 314, 413–418.

    Google Scholar 

  • Löhr, F., Blümel, M., Schmidt, J.M. and Rüterjans, H. (1997) J. Biomol. NMR, 10, 107–118.

    Google Scholar 

  • Löhr, F. and Rüterjans, H. (1995) J. Biomol. NMR, 5, 25–36.

    Google Scholar 

  • Löhr, F. and Rüterjans, H. (1999) J. Biomol. NMR, 13, 263–274.

    Google Scholar 

  • Marino, J.P., Diener, J.L., Moore, P.B. and Griesinger, C. (1997) J. Am. Chem. Soc., 85, 2870–2871.

    Google Scholar 

  • Marion, D., Ikura, M., Tschudin, R. and Bax, A. (1989) J. Magn. Reson., 85, 393–399.

    Google Scholar 

  • Martinez-Oyanedel, J., Choe, H.W., Heinemann U. and Saenger, W. (1991) J. Mol. Biol., 222, 335–352.

    Google Scholar 

  • Mattiello, D.L., Warren, W.S., Mueller, L. and Farmer II, B.T. (1996) J. Am. Chem. Soc., 118, 3253–3261.

    Google Scholar 

  • Norwood, T.J. (1992) Prog. NMR Spectrosc., 24, 295–375.

    Google Scholar 

  • Piantini, U., Sørensen, O.W. and Ernst, R.R. (1982) J. Am. Chem. Soc., 104, 6800–6801.

    Google Scholar 

  • Schmidt, J.M. (1997) J. Magn. Reson., 124, 298–309.

    Google Scholar 

  • Schmidt, J.M., Blümel, M., Löhr, F. and Rüterjans, H. (1999) J. Biomol. NMR, 14, 1–12.

    Google Scholar 

  • Schmidt, J.M., Ernst, R.R., Aimoto, S. and Kainosho, M. (1995) J. Biomol. NMR, 6, 95–105.

    Google Scholar 

  • Schmidt, J.M., Löhr, F. and Rüterjans, H. (1996) J. Biomol. NMR, 7, 142–152.

    Google Scholar 

  • Schmidt, J.M. and Rüterjans, H. (1990) J. Am. Chem. Soc., 112, 1279–1280.

    Google Scholar 

  • Seip, S., Balbach, J. and Kessler, H. (1992) J. Magn. Reson., 100, 406–410.

    Google Scholar 

  • Shaka, A.J., Barker, P.B. and Freeman, R. (1985) J. Magn. Reson., 64, 547–552.

    Google Scholar 

  • Sklenár, V., Dieckmann, T., Butcher, S.E. and Feigon, J. (1998) J. Magn. Reson., 130, 119–124.

    Google Scholar 

  • Smallcombe, S.H., Patt, S.L. and Keifer, P.A. (1995) J. Magn. Reson., A117, 295–303.

    Google Scholar 

  • Swapna, G.V.T., Rios, C.B., Shang, Z. and Montelione, G.T. (1997) J. Biomol. NMR, 9, 105–111.

    Google Scholar 

  • Tessari, M., Gentile, L.N., Taylor, S.J., Shalloway, D.I., Nicholson, L.K. and Vuister, G.W. (1997) Biochemistry, 36, 14561–14571.

    Google Scholar 

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

    Google Scholar 

  • Wasylishen, R. and Schaefer, T. (1972) Can. J. Chem., 50, 2710–2712.

    Google Scholar 

  • Watt, W., Tulinsky, A., Swenson, R.P. and Watenpaugh, K.D. (1991) J. Mol. Biol., 218, 195–208.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Biophysikalische Chemie, Johann Wolfgang Goethe-Universität, Biozentrum N230, Marie-Curie-Strasse 9, D-60439, Frankfurt am Main, Germany

    Frank Löhr, Carlos Pérez, Rolf Köhler & Heinz Rüterjans

  2. Division of Molecular Structure, National Institute for Medical Research, Mill Hill, London, NW7 1AA, U.K.

    Jürgen M. Schmidt

Authors
  1. Frank Löhr
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carlos Pérez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rolf Köhler
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heinz Rüterjans
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jürgen M. Schmidt
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Löhr, F., Pérez, C., Köhler, R. et al. Heteronuclear relayed E.COSY revisited: Determination of 3J(Hα,Cγ) couplings in Asx and aromatic residues in proteins. J Biomol NMR 18, 13–22 (2000). https://doi.org/10.1023/A:1008385202639

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008385202639

Search

Navigation