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Journal of Biomolecular NMR

, Volume 4, Issue 6, pp 787–797 | Cite as

Resonance assignment of methionine methyl groups and χ3angular information from long-range proton—carbon and carbon—carbon J correlation in a calmodulin—peptide complex

  • Ad Bax
  • Frank Delaglio
  • Stephan Grzesiek
  • Geerten W. Vuister
Research Paper

Summary

Several simple 3D experiments are used to provide J correlations between methionine Cε methyl carbons and either the CγH2 protons or Cβ and Cγ. The intensity of the J correlations provides information on the size of the three-bond J couplings and thereby on the χ3 torsion angle. In addition, a simple 3D version of the HMBC experiment provides a sensitive link between the CεH3 methyl protons and Cγ. The methods are demonstrated for a 20 kDa complex between calmodulin and a 26-residue peptide fragment of skeletal muscle myosin light chain kinase.

Keywords

Calmodulin Carbon-carbon J coupling Carbon-proton J coupling HMBC Methionine Protein NMR 3D NMR Isotope labeling 

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References

  1. BaxA. and PochapskyS.S. (1992) J. Magn. Reson., 99, 638–643.Google Scholar
  2. BaxA. and SummersM.F. (1986) J. Am. Chem. Soc., 108, 2093–2094.Google Scholar
  3. BaxA., CloreG.M. and GronenbornA.M. (1990) J. Magn. Reson., 88, 425–431.Google Scholar
  4. BaxA., MaxD. and ZaxD. (1992) J. Am. Chem. Soc., 114, 6924–6925.Google Scholar
  5. CohenP. and KleeC.B. (1988) Molecular Aspects of Cellular Regulation, Vol. 5, Elsevier, New York, NY.Google Scholar
  6. FesikS.W., EatonH.L., OlejniczakE.T., ZuiderwegE.R.P., McIntoshL.P. and DahlquistF.W. (1990) J. Am. Chem. Soc., 112, 886–888.Google Scholar
  7. FinnB.E., DrakenbergT. and ForsenS. (1994) FEBS Lett., 336, 368–374.Google Scholar
  8. GrzesiekS. and BaxA. (1992) J. Magn. Reson., 99, 201–207.Google Scholar
  9. IkuraM., CloreG.M., GronenbornA.M., ZhuG., KleeC.B. and BaxA. (1992) Science, 256, 632–638.Google Scholar
  10. KayL.E., IkuraM. and BaxA. (1990) J. Am. Chem. Soc., 112, 888–889.Google Scholar
  11. KlevitR.E., DalgarnoD.C., LevineB.A. and WilliamsR.J.P. (1984) Biochemistry, 24, 8152–8157.Google Scholar
  12. KretsingerR. (1992) Cell Calcium, 13, 363–376.Google Scholar
  13. KrivdinI.B. and DellaE.W. (1991) Prog. NMR Spectrosc., 23, 301–610.Google Scholar
  14. MarionD., IkuraM., TschudinR. and BaxA. (1989) J. Magn. Reson., 85, 393–399.Google Scholar
  15. MeadorW.E., MeansA.R. and QuiochoF.A. (1992) Science, 257, 1251–1255.Google Scholar
  16. MeadorW.E., MeansA.R. and QuiochoF.A. (1993) Science, 262, 1718–1721.Google Scholar
  17. O'NeilK.T. and DeGradoW.F. (1990) Trends Biochem. Sci, 15, 59–64.Google Scholar
  18. SantoroJ. and KingG.C. (1992) J. Magn. Reson., 97, 202–207.Google Scholar
  19. Van deVenF.J.M. and PhilippensM.E.P. (1992) J. Magn. Reson., 97, 637–644.Google Scholar
  20. VuisterG.W. and BaxA. (1992) J. Magn. Reson., 98, 428–435.Google Scholar
  21. VuisterG.W. and BaxA. (1993) J. Magn. Reson. Ser. B, 102, 228–231.Google Scholar
  22. VuisterG.W., YamazakiT., TorchiaD.A. and BaxA. (1993) J. Biomol. NMR, 3, 297–306.Google Scholar
  23. VuisterG.W., GrzesiekS., DelaglioF., WangA.C., TschudinR., ZhuG. and BaxA. (1994) Methods Enzymol., 239, 79–105.Google Scholar
  24. ZhuG. and BaxA. (1990) J. Magn. Reson., 90, 405–410.Google Scholar
  25. ZhuG. and BaxA. (1993) J. Magn. Reson. Ser. A., 104, 353–357.Google Scholar

Copyright information

© ESCOM Science Publishers B.V 1994

Authors and Affiliations

  • Ad Bax
    • 1
  • Frank Delaglio
    • 1
  • Stephan Grzesiek
    • 1
  • Geerten W. Vuister
    • 1
  1. 1.Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaU.S.A.

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