Journal of Biomolecular NMR

, Volume 4, Issue 6, pp 761–774 | Cite as

113Cd-1H hetero TOCSY: A method for determining metal—protein connectivities

  • Kevin H. Gardner
  • Joseph E. Coleman
Research Paper


113Cd-1H NMR correlation experiments have been extremely useful for determining the amino acid ligands that form metal-binding sites in proteins. To date, the majority of these methods have used heteronuclear multiple-quantum transfer as the basis for establishing correlations. In this paper, we demonstrate the feasibility of using correlation methods that employ heteronuclear cross-polarization (hetero TOCSY) as viable alternatives. Additionally, we couple hetero TOCSY with selective excitation and transfer procedures to take advantage of the small number of heteronuclei usually present in metalloprotein systems. One- and two-dimensional experiments are presented as examples of these techniques.


NMR Metalloproteins 113Cd Heteronuclear cross-polarization Hetero TOCSY 


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  1. BalejaJ.D., MarmorsteinR., HarrisonS.C., and WagnerG. (1992) Nature, 356, 450–453.Google Scholar
  2. BeardenD.W. and BrownL.R. (1989) Chem. Phys. Lett., 163, 432–436.Google Scholar
  3. BlakeP.R., LeeB., SummersM.F., ParkJ.-B., ZhouZ.H., and AdamsM.W.W. (1994) New J. Chem., 18, 387–395.Google Scholar
  4. BoudotD., CanetD., BrondeauJ. and BoubelJ.C. (1989) J. Magn. Reson., 83, 428–439.Google Scholar
  5. BrownL.R., and SanctuaryB.C. (1991) J. Magn. Reson., 91, 413–421.Google Scholar
  6. ColemanJ.E. (1993) Methods Enzymol., 227, 16–43.Google Scholar
  7. ErnstM., GriesingerC., ErnstR.R. and BermelW. (1991) Mol. Phys., 74, 219–252.Google Scholar
  8. FreyM.H., WagnerG., VasákM., SørensenO.W., NeuhausD.G., WörgötterE., KägiJ.H.R., ErnstR.R. and WüthrichK. (1985) J. Am. Chem. Soc., 107 6847–6851.Google Scholar
  9. GadhaviP.L., RaineA.R.C., AlefounderP.R. and LaueE.D., (1990) FEBS Lett. 276, 49–53.Google Scholar
  10. GadhaviP.L., DavisA.L., PoveyJ.F., KeelerJ. and LaueE.D. (1991) FEBS Lett., 281, 223–226.Google Scholar
  11. GardnerK.H., PanT., NarulaS., RiveraE., ColemanJ.E. (1991) Biochemistry, 30, 11292–11302.Google Scholar
  12. GlaserS.J. and DrobnyG.P. (1989) Chem. Phys. Lett., 164, 456–462.Google Scholar
  13. GlaserS.J. and DrobnyG.P. (1991) Chem. Phys. Lett., 184, 553–559.Google Scholar
  14. HenehanC.J., PountneyD.L., ZerbeO. and VasákM. (1993) Protein Sci., 2, 1756–1764.Google Scholar
  15. KayL.E., MarionD. and BaxA. (1989) J. Magn. Reson., 84, 72–84.Google Scholar
  16. KelloggG.W. (1992) J. Magn. Reson., 98, 176–182.Google Scholar
  17. KelloggG.W. and SchweitzerB.I. (1993) J. Biomol. NMR, 3, 577–595.Google Scholar
  18. KesslerH., OschkinatH., GriesingerC. and BermelW. (1986) J. Magn. Reson., 70, 106–133.Google Scholar
  19. KraulisP.J. (1991) J. Appl. Cryst., 24, 946–950.Google Scholar
  20. KraulisP.J., RaineA.R.C., GadhaviP.L. and LaueE.D. (1992) Nature, 356, 448–450Google Scholar
  21. MajumdarA., WangH., MorshauserC., and ZuiderwegE.R.P. (1993) J. Biomol. NMR, 3, 387–397.Google Scholar
  22. MarmorsteinR., CareyM., PtashneM. and HarrisonS.C. (1992) Nature, 356, 408–414Google Scholar
  23. MüllerL. and ErnstR.R. (1979) Mol. Phys., 38, 963–992.Google Scholar
  24. OttingG., SennH., WagnerG. and WüthrichK. (1986) J. Magn. Reson., 70, 500–505.Google Scholar
  25. PanT. and ColemanJ.E. (1989) Proc. Natl. Acad. Sci. USA, 86, 3145–3149.Google Scholar
  26. PanT. and ColemanJ.E. (1990a) Biochemistry, 29, 3023–3029.Google Scholar
  27. PanT. and ColemanJ.E. (1990b) Proc. Natl. Acad. Sci. USA, 87, 2077–2081.Google Scholar
  28. PanT., HalvorsenY.-D., DicksonR.C. and ColemanJ.E. (1990) J. Biol. Chem. 265, 21427–21429.Google Scholar
  29. RanceM. (1987) J. Magn. Reson., 74, 557–564.Google Scholar
  30. RichardsonJ.M., ClowesR.T., BoucherW., DomailleP.J., HardmanC.H., KeelerJ. and LaueE.D. (1993) J. Magn. Reson. Ser. B, 101, 223–227.Google Scholar
  31. SchmidtP., SchwalbeH., GlaserS.J. and GriesingerC. (1993) J. Magn. Reson. Ser. B, 101, 328–332.Google Scholar
  32. ShakaA.J., LeeC.J. and PinesA. (1988) J. Magn. Reson., 77, 274–293.Google Scholar
  33. ShirakawaM., FairbrotherW.J., SerikawaY., OhkuboT., KyogokuY. and WrightP.E. (1993) Biochemistry, 32, 2144–2153.Google Scholar
  34. SørensenO.W. and ErnstR.R. (1983) J. Magn. Reson., 51, 477–489.Google Scholar
  35. SørensenO.W., RanceM. and ErnstR.R. (1984) J. Magn. Reson., 56, 527–534.Google Scholar
  36. WangK.Y., GoljerI. and BoltonP.H. (1994) J. Magn. Reson. Ser. B, 103, 192–196.Google Scholar
  37. ZerbeO., PountneyD.L., VonPhilipsbornW. and VasákM. (1994) J. Am. Chem. Soc., 116, 377–378.Google Scholar
  38. ZuiderwegE.R.P. (1990) J. Magn. Reson., 89, 533–542.Google Scholar

Copyright information

© ESCOM Science Publishers B.V 1994

Authors and Affiliations

  • Kevin H. Gardner
    • 1
  • Joseph E. Coleman
    • 1
  1. 1.Department of Molecular Biophysics and BiochemistryYale UniversityNew HavenU.S.A.

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