Three- and Four-Dimensional Heteronuclear NMR

  • G. Marius Clore
  • Angela M. Gronenborn


The principal source of geometric information used to solve three dimensional structures of macromolecules by NMR resides in short (< 5Å) approximate interproton distance restraints derived from nuclear Overhauser enhancement (NOE) measurements (1–5). In order to extract this information it is essential to first completely assign the 1H spectrum of the macromolecule in question and then to assign as many structurally useful NOE interactions as possible. The larger the number of NOE restraints, the higher the precision and accuracy of the resulting structures (5–7). Indeed, with current state-of-the-art methodology it is now possible to obtain NMR structures of proteins at a precision and accuracy comparable to 2 Å resolution crystal structures (7–9)


Cross Peak NOESY Spectrum Pulse Field Gradient Pulse Scheme Nuclear Overhauser Enhancement 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Wüthrich, K., 1986, NMR of Proteins, Wiley, New YorkGoogle Scholar
  2. 2.
    Clore, G.M., and Gronenborn, A.M., 1989, Determination of three-dimensional structures of proteins in solution by nuclear magnetic resonance spectroscopy. Prot. Eng. 1: 275–288.Google Scholar
  3. 3.
    Clore, G.M., and Gronenborn, A.M., 1989, Determination of three-dimensional structures of proteins and nucleic acids in solution by nuclear magnetic resonance spectroscopy. CRC Crit Rev. Biochem. Mol. Biol. 24: 479–564.CrossRefGoogle Scholar
  4. 4.
    Bax, A., 1989, Two-dimensional NMR and protein structure, Ann Rev. Biochem. 58: 223–256.PubMedCrossRefGoogle Scholar
  5. 5.
    Clore, G.M., and Gronenborn, A.M., 1991, Structures of larger proteins in solution: three-and four-dimensional hetronuclear NMR spectroscopy. Science 252: 1390–1399.PubMedCrossRefGoogle Scholar
  6. 6.
    Clore, G.M., and Gronenborn, A.M., 1991, Two, three and four dimensional NMR methods for obtaining larger and more precise three-dimensional structures of proteins in solution. Ann. Rev. Biophys. Biophys. Chem. 20: 29–63.CrossRefGoogle Scholar
  7. 7.
    G.M. Clore, and A.M. Gronenborn, 1991, Comparison of the solution nuclear magnetic resonance and X-ray crystal structures of human recombinant interleukin-113. J. Mol. Biol. 221: 47–53.PubMedCrossRefGoogle Scholar
  8. 8.
    Clore, G.M., Robien, M.A., and Gronenborn, A.M., 1993, Exploring the limits of precision and accuracy of protein structures determined by nuclear magnetic resonance spectroscopy. J. Mol. Biol. 231: 82–102.PubMedCrossRefGoogle Scholar
  9. 9.
    Shaanan, B., Gronenborn, A.M., Cohen, G.H., Gilliland, G.L., Veerapandian, B., Davies, D.R., and Clore, G.M., 1992, Combining experimental information from crystal and solution studies: joint X-ray and NMR refinement, Science 257: 961–964.PubMedCrossRefGoogle Scholar
  10. 10.
    Dyson, H.J., Gippert, D.A., Case, D.A., Holmgren, A., and Wright, P.E., 1990, Three-dimensional solution structure of the reduced form of Escherichia coli thioredoxin determined by nuclear magnetic resonance spectroscopy, Biochemistry 29: 4129–4136.PubMedCrossRefGoogle Scholar
  11. 11.
    Forman-Kay, J.D., Clore, G.M., Wingfield, P.T., and Gronenborn, A.M., 1991, The high resolution three-dimensional structure of reduced recombinant human thioredoxin in solution, Biochemistry 30: 2685–2698.PubMedCrossRefGoogle Scholar
  12. 12.
    Oschkinat, H., Griesinger, C., Kraulis, P.J., Sorensen, O.W., Ernst, R.R., Gronenbor, A.M., and Clore, G.M., 1988, Three-dimensional NMR spectroscopy of a protein in solution, Nature (Lond.) 332: 374–376.CrossRefGoogle Scholar
  13. 13.
    Clore, G.M., and Gronenborn, A.M., 1991, Applications of three-and four-dimensional heteronuclear NMR spectroscopy to protein structure determination, Progr. Nucl. Magn. Reson. Spectrosc. 23: 43–92.CrossRefGoogle Scholar
  14. 14.
    Bax, A., and Grzesiek, S., 1993, Methodological Advances in Protein NMR, Acct. Chem. Res. 26: 131–138.CrossRefGoogle Scholar
  15. 15.
    Kay, L.E., Clore, G.M., Bax, A., and Gronenborn, A.M., 1990, Four-dimensional heteronuclear triple resonance NMR spectroscopy of interleukin-113 in solution, Science 249: 411–414.PubMedCrossRefGoogle Scholar
  16. 16.
    Clore, G.M., Kay, L.E., Bax, A., and Gronenborn, A.M., 1991, Four-dimensional 13C/13C-edited nuclear Overhauser enhancement spectroscopy of a protein in solution: application to interleukin-10. Biochemistry 30, 12–18.PubMedCrossRefGoogle Scholar
  17. 17.
    Zuiderweg, E.R.P., Petros, A.M., Fesik, S.W., and Olejniczak, E.T., 1991, Four-dimensional [13C, 1H, 13C, H] HMQC-NOE-HMQC NMR spectroscopy: resolving tertiary NOE distance restraints in spectra of larger proteins, J. Am. Chem. Soc. 113: 370–372.CrossRefGoogle Scholar
  18. 18.
    Bax, A., Clore, G.M., Driscoll, P.C., Gronenborn, A.M., Ikura, M., and Kay, L.E., 1990, Practical aspects of proton-carbon-carbon-proton three-dimensional correlation spectroscopy of 13C-labeled proteins. J. Magn. Reson. 87: 620–628.Google Scholar
  19. 19.
    Bax, A., Clore, G.M., and Gronenborn, A.M., 1990, 1H–1H correlation via isotropic mixing of 13C magnetization: a new three-dimensional approach for assigning 1H and 13C spectra of 13C-enriched proteins. J. Magn. Reson. 88, 425–431.Google Scholar
  20. 20.
    Fesik, S.W., Eaton, H.L., Olejniczak, E.T., Zuiderweg, E.R.P., McIntosh, L.P., and Dahlquist, F.W., 1990, 2D and 3D NMR spectroscopy employing 13C–13C magentization transfer by isotropic mixing: spin system identification in large proteins, J. Am. Chem. Soc. 112, 886–888.Google Scholar
  21. 21.
    Clore, G.M., Bax, A., Driscoll, P.C., Wingfield, P.T., and Gronenborn, A.M., 1990, Assignment of the side chain IH and 13C resonances of interleukin-13 using double and triple resonance hetronuclear three-dimensional NMR spectroscopy. Biochemistry 29: 8172–8184.PubMedCrossRefGoogle Scholar
  22. 22.
    Ikura, M., Kay, L.E., and Bax, A., 1990, A novel approach for sequential assignment of ‘H, 13C, and 15N spectra of larger protens: heteronuclear triple-resonance NMR spectroscopy: application to calmodulin, Biochemistry 29, 4659–4667.PubMedCrossRefGoogle Scholar
  23. 23.
    Powers, R., Gronenborn, A.M., Clore, G.M., and Bax, A., 1991, Three-dimensional triple resonance NMR of 13C/15N enriched proteins using constant-time evolution. J. Magn. Reson. 94: 209–213.Google Scholar
  24. 24.
    Grzesiek, S., and Bax, A., 1992, Correlating backbone amide and sidechain resonances in larger proteins by multiple relayed triple resonance NMR, J. Am. Chem. Soc. 114: 6291–6293.CrossRefGoogle Scholar
  25. 25.
    Grzesiek, S., and Bax, A., 1993, Amino acid type determination in the sequential assignment procedure of uniformly 13C/15N enriched proteins, J. Biomol. NMR 3: 185–204.PubMedGoogle Scholar
  26. 26.
    Grzesiek, S., Anglister, J., and Bax, A., 1993, Correlation of backbone amide and aliphatic sidechain resonances in 13C/15N-enriched proteins by isotropic mixing of 13C magnetization, J. Magn. Reson. Series B 101: 114–119.CrossRefGoogle Scholar
  27. 27.
    Grzesiek, S., and Bax, A., 1992, An efficient experiment for sequential backbone assignment of medium sized isotopically enriched proteins, J. Magn. Reson. 99: 201–207.Google Scholar
  28. 28.
    Clore, G.M., Bax, A., and Gronenborn, A.M., 1991, Stereospecific assignment of b-methylene protons in larger proteins using three-dimensional 15N-separated Hartmann-Hahn and 13C-separated rotating frame Overhauser spectroscopy. J. Biomol. NMR 11: 13–22.CrossRefGoogle Scholar
  29. 29.
    Bax, A., and Pochapsky, S.J., 1992, Optimized recording of heteronuclear multi-dimensional NMR spectra using pulsed field gradients, J. Magn. Reson. 99: 638–643.Google Scholar
  30. 30.
    Vuister, G.W., Clore, G.M., Gronenborn, A.M., Powers, R., Garrett, D.S., Tschudin, R, and Bax, A., 1993, Increased resolution and improved spectral quality in four-dimensional 13C/13C separated HMQC-NOEHMQC spectra using pulsed field gradients, J. Magn. Reson. Series B 101: 210-–213.Google Scholar
  31. 31.
    Clore, G.M., Wingfield, P.T., and Gronenbom, A.M., 1991, High resolution three-dimensional structure of interleukin-113 in solution by three and four dimensional nuclear magnetic resonance spectroscopy. Biochemistry 30: 2315–2323.PubMedCrossRefGoogle Scholar
  32. 32.
    Powers, R., Garrett, D.S., March, C.J., Frieden, E.A., Gronenborn, A.M., and Clore, G.M., 1992, Three-dimensional solution structure of interleukin-4 by multi-dimensional heteronuclear magnetic resonance spectroscopy, Science 256: 1673–1677.PubMedCrossRefGoogle Scholar
  33. 33.
    Smith, L.J., Redfield, C., Boyd, J., Lawrence, G.M.P., Edwards, R.G., Smith, R.A.G., and Dobson, C.M., 1992, Human interleukin-4: the solution structure of a four helix bundle protein. J. Mol. Biol. 224: 900–904.CrossRefGoogle Scholar
  34. 34.
    Powers, R., Garrett, D.S., March, C.J., Frieden, E.A., Gronenbom, A.M., and Clore, G.M., 1993, The high resolution three-dimensional solution structure of interleukin-4 determined by multi-dimensional heteronuclear magnetic resonance spectroscopy, Biochemistry 32: 6744–6762.PubMedCrossRefGoogle Scholar
  35. 35.
    Fairbrother, W.J., Gippert, G.P., Reizer, J., Saier, M.J., and Wright, P.E., 1992, Low resolution structure of the Bacillus subtilis glucose permease IIA domain derived from heteronuclear three-dimensional NMR spectroscopy, FEBS Lett. 296: 148–152.PubMedCrossRefGoogle Scholar
  36. 36.
    Ikura, M., Clore, G.M., Gronenborn, A.M., Zhu, G., Klee, C.B., and Bax, A., 1992, Solution structure of a calmodulin-target peptide complex by multi-dimensional NMR, Science 256: 632–638.PubMedCrossRefGoogle Scholar
  37. 37.
    Thierault, Y., Logan, T.M., Meadows, R., Yu, L., Olejniczak, E.T., Holzman, T.F., Sikmmer, R.L., and fesik, S.W., 1993, Solution structure of the cyclosporin A/cyclophilin complex by NMR, Nature (Lond.) 361, 88–91.CrossRefGoogle Scholar
  38. 38.
    Omichinski, J.G., Clore, G.M., Schaad, O., Felsenfeld, G., Trainor, C., Appella, E., Stahl, S.J., and Gronenborn, A.M., 1993, NMR structure of a specific DNA complex of Zn-containing DNA binding domain of GATA-1, Science 261: 438–446.PubMedCrossRefGoogle Scholar
  39. 39.
    Lodi, P.J., Garrett, D.S., Kuszewski, J., Tsang, M.L.S., Weatherbee, J.A., Leonard, W.J., Gronenborn, A.M., and Clore, G.M., 1994, High resolution solution structure of the ß chemokine hMIP-113 by multi-dimensional NMR, Science 263: 1762–1767.PubMedCrossRefGoogle Scholar
  40. 40.
    Clore, G.M., Omichinski, J.G., Sakaguchi, K., Zambrano, N., Sakamoto, H., Appella, E., and Gronenbom, A.M., 1994, High-resolution solution structure of the oligomerization domain of p53 by multi-dimensional NMR, Science 265: 386–391.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • G. Marius Clore
    • 1
  • Angela M. Gronenborn
    • 1
  1. 1.Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA

Personalised recommendations