NMR Studies of Protein-Ligand Interactions: Dihydrofolate Reductase

  • G. C. K. Roberts
Part of the NATO ASI Series book series (NSSA, volume 107)


The study of protein-ligand interactions has proved to be one of the more fruitful applications of high-resolution nmr to biological problems. The specific recognition of small molecules by proteins is fundamental to a wide range of biological phenomena, from enzyme action and its regulation, through the control of gene expression to the action of hormones and neurotransmitters. Nmr can provide a variety of information on both structural and dynamic aspects of the recognition process in the solution state. In this article I shall describe some of the different kinds of Information which can be obtained, drawing illustrations from work on the enzyme dlhydrofolate reductase, which we have been studying for a number of years. A broader view of the wide range of experiments which have been done on other systems can be found in ref. 1, while fuller references to the work on dlhydrofolate reductase are given in ref. 2. Operationally, two kinds of experiments can be distinguished, depending on whether one studies resonances of the small molecule or of the protein, and examples of both will be discussed. First, however, we must consider, briefly, the effects of chemical exchange processes on nmr spectra and their interpretation.


Chemical Shift Free State Ligand Concentration Dihydrofolate Reductase Dissociation Rate Constant 
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  1. 1.
    Jardetzky, O. and Roberts, G.C.K. (1981). Nmr in Molecular Biology. Academic Press, New York.Google Scholar
  2. 2.
    Feeney, J. (1985) in “Nmr in Living Systems” (Proc. NATO ASI) D. Reidel, Dordrecht, in press.Google Scholar
  3. 3.
    Birdsall, B., Roberts, G.C.K., Feeney, J. and Burgen, A.S.V. (1977) FEBS Lett., 80:313.PubMedCrossRefGoogle Scholar
  4. 4.
    Hyde, E.I., Birdsall, B., Roberts, G.C.K., Feeney, J. and Burgen, A.S.V. (1980) Biochemistry, 19:3746.PubMedCrossRefGoogle Scholar
  5. 5.
    Feeney, J., Batchelor, J.G., Albrand, J.P. and Roberts, G.C.K. (1979) J. Mag. Reson., 33:519.CrossRefGoogle Scholar
  6. 6.
    Gronenborn, A.M., Birdsall, B., Hyde, E.I., Roberts, G.C.K., Feeney, J. and Burgen, A.S.V. (1981) Molec. Pharmacol., 20:145.Google Scholar
  7. 7.
    Birdsall, B., Gronenborn, A., Hyde, E.I., Clore, G.M., Roberts, G.C.K., Feeney, J. and Burgen, A.S.V. (1982) Biochemistry, 21:5831.PubMedCrossRefGoogle Scholar
  8. 8.
    Birdsall, B., Bevan, A.W., Pascual, C., Roberts, G.C.K., Feeney, J., Gronenborn, A. and Clore, G.M. (1984) Biochemistry, 23:4733.PubMedCrossRefGoogle Scholar
  9. 9.
    Birdsall, B., Hammond, S., De Graw, J.I., Roberts, G.C.K. and Feeney, J. (1986) Submitted for publication.Google Scholar
  10. 10.
    Forsen, S. and Hoffman, R.A. (1963) J. Chem. Phys., 39:2892.CrossRefGoogle Scholar
  11. 11.
    Hyde, E.I., Birdsall, B., Roberts, G.C.K., Feeney, J. and Burgen, A.S.V. (1980) Biochemistry, 19:3738.PubMedCrossRefGoogle Scholar
  12. 12.
    Clore, G.M., Roberts, G.C.K., Gronenborn, A., Birdsall, B. and Feeney, J. (1981) J. Mag. Reson., 45:141.Google Scholar
  13. 13.
    Birdsall, B., Hyde, E.I., Burgen, A.S.V., Roberts, G.C.K. and Feeney, J. (1981) Biochemistry, 19:3732.CrossRefGoogle Scholar
  14. 14.
    Birdsall, B., Roberts, G.C.K., Feeney, J., Dann, J.G. and Burgen, A.S.V. (1983) Biochemistry, 22:5597.PubMedCrossRefGoogle Scholar
  15. 15.
    Albrand, J.P., Birdsall, B., Feeney, J., Roberts, G.C.K. and Burgen, A.S.V. (1979) Int. J. Biol. Macromol., 1:37.CrossRefGoogle Scholar
  16. 16.
    Clore, C.M. and Gronenborn, A.M. (1982) J. Mag. Reson., 48:402.CrossRefGoogle Scholar
  17. 17.
    Feeney, J., Birdsall, B., Roberts, G.C.K. and Burgen, A.S.V. (1983) Biochemistry, 22:628.PubMedCrossRefGoogle Scholar
  18. 18.
    Baker, D.J., Beddell, C.R., Champness, J.N., Goodford, P.J., Norrington, F.E.A., Smith, D.R. and Stammers, D.K. (1981) FEBS Lett., 126:49.PubMedCrossRefGoogle Scholar
  19. 19.
    Roberts, G.C.K., Feeney, J., Burgen, A.S.V. and Daluge, S. (1981) FEBS Lett., 131:65.CrossRefGoogle Scholar
  20. 20.
    Morris, G.A. and Freeman, R. (1979) J. Am. Chem. Soc., 101:760.CrossRefGoogle Scholar
  21. 21.
    Morris, G.A. (1980) J. Am. Chem. Soc., 102:428.CrossRefGoogle Scholar
  22. 22.
    Bevan, A.W., Roberts, G.C.K., Feeney, J. and Kuyper, L. (1985) Eur. Biophys. J., 11:21.CrossRefGoogle Scholar
  23. 23.
    Cayley, P.J., Albrand, J.P., Feeney, J., Roberts, G.C.K., Piper, E.A. and Burgen, A.S.V. (1979) Biochemistry, 18:3886.PubMedCrossRefGoogle Scholar
  24. 24.
    Campbell, I.D., Dobson, C.M. and Williams, R.J.P. (1975), Proc. Roy. Soc. B., 189:503.CrossRefGoogle Scholar
  25. 25.
    Wuthrich, K. and Wagner, G. (1975) FEBS Lett., 50:265.PubMedCrossRefGoogle Scholar
  26. 26.
    Karplus, M. and McCammon, J.A. (1981) CRC Crit. Rev. Biochem., 9:293.PubMedCrossRefGoogle Scholar
  27. 27.
    Feeney, J., Birdsall, B., Albrand, J.P., Roberts, G.C.K., Burgen, A.S.V., Charlton, P.A. and Young, D.W. (1981) Biochemistry, 20:1837.PubMedCrossRefGoogle Scholar
  28. 28.
    Clore, C.M., Gronenborn, A.M., Birdsall, B., Feeney, J. and Roberts, G.C.K. (1984), Biochem. J., 217:659.PubMedGoogle Scholar
  29. 29.
    Cheung, H.T.A., Searle, M.S., Feeney, J., Birdsall, B., Roberts, G.C.K., Kompis, I. and Hammond, S. (1986), Biochemistry, in press.Google Scholar
  30. 30.
    Wuthrich, K., Wider, G., Wagner, G. and Braun, W. (1982), J. Mol. Biol., 155:311.PubMedCrossRefGoogle Scholar
  31. 31.
    Billeter, M., Braun, W. and Wuthrich, K. (1982), J. Mol. Biol., 155:321.PubMedCrossRefGoogle Scholar
  32. 32.
    Wagner, G. and Wuthrich, K. (1982) J. Mol. Biol., 155:347.PubMedCrossRefGoogle Scholar
  33. 33.
    Strop, P., Wider, G. and Wuthrich, K. (1983), J. Mol. Biol., 166:641.PubMedCrossRefGoogle Scholar
  34. 34.
    Dobson, C.M., Howarth, M.A. and Redfield, C. (1984), FEBS Lett., 176:307.CrossRefGoogle Scholar
  35. 35.
    Delepierre, M., Dobson, C.M., Howarth, M.A. and Poulsen, F.M. (1984) Eur. J. Biochem., 145:389.PubMedCrossRefGoogle Scholar
  36. 36.
    Aue, W.P., Bartholdi, E. and Ernst, R.R. (1976), J. Chem. Phys., 64:2229.CrossRefGoogle Scholar
  37. 37.
    Eich, G., Bodenhausen, G. and Ernst, R.R. (1982) J. Am. Chem. Soc., 104:3732.CrossRefGoogle Scholar
  38. 38.
    Feeney, J., Roberts, G.C.K., Birdsall, B., Griffiths, D.V., King, R.W., Scudder, P. and Burgen, A.S.V. (1971), Proc. Roy. Soc. B., 196:267.CrossRefGoogle Scholar
  39. 39.
    Feeney, J., Roberts, G.C.K., Thomson, J., King, R.W., Griffiths, D.V. and Burgen, A.S.V. (1980), Biochemistry, 19:2316.PubMedCrossRefGoogle Scholar
  40. 40.
    Birdsall, B., Feeney, J., Griffiths, D.V., Hammond, S., Kimber, B., King, R.W., Roberts, G.C.K. and Searle, M.S. (1984), FEBS Lett., 175:364.PubMedCrossRefGoogle Scholar
  41. 41.
    Williamson, M.P., Marion, D. and Wuthrich, K. (1984), J. Mol. Biol., 173:341.PubMedCrossRefGoogle Scholar
  42. 42.
    Hammond, S.J., Birdsall, B., Searle, M.S., Roberts, G.C.K. and Feeney, J. (1986), J. Mol. Biol., in press.Google Scholar
  43. 43.
    Antonjuk, D.J., Birdsall, B., Burgen, A.S.V., Cheung, H.T.A., Clore, G.M., Feeney, J., Gronenborn, A., Roberts, G.C.K. and Tran, T.Q. (1984), Brit. J. Pharmacol., 81:309.Google Scholar
  44. 44.
    Birdsall, B., Feeney, J., Pascual, C., Roberts, G.C.K., Kompis, I., Then, R.L., Muller, K. and Kroehn, A. (1984), J. Med. Chem., 27:1672.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • G. C. K. Roberts
    • 1
  1. 1.Division of Physical BiochemistryNational Institute for Medical ResearchMill Hill, LondonUK

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