Journal of Biomolecular NMR

, Volume 10, Issue 3, pp 289–292 | Cite as

High-resolution heteronuclear NMR of human ubiquitin in an aqueous liquid crystalline medium

  • Ad Bax
  • Nico Tjandra

Abstract

A mixture of dihexanoyl phosphatidylcholine and dimyristoylphosphatidylcholine in water forms disc-shaped particles, often referred toas bicelles [Sanders and Schwonek (1992) Biochemistry, 31, 8898–8905].These adopt an ordered, liquid crystalline phase, which can be maintained atvery low concentrations of the bicelles (down to 3% w/v). At thisconcentration the spacing between individual bicelles, on average, exceeds300 Å. The bicelles are shown to have a negligible effect on therotational diffusion of ubiquitin as judged by the 15NT values of the backbone amides relative to those inisotropic aqueous solution. The protein exhibits a residual degree ofalignment which is proportional to the bicelle concentration, andapproximately collinear with ubiquitin's rotational diffusion tensor. Thedegree of alignment obtained offers unique opportunities for studying theprotein's structure and dynamics.

Alignment Bicelle Dipolar coupling Lipid Liquid crystal 15N relaxation 

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References

  1. Cavanagh, J., Fairbrother, W., Palmer III, A.G. and Skelton, N.J. (1995) Protein NMR Spectroscopy, Academic Press, San Diego, CA, U.S.A.Google Scholar
  2. Clore, G.M. and Gronenborn, A.M. (1989) CRC Crit. Rev. Biochem. Mol. Biol., 24, 479–564.Google Scholar
  3. Diehl, P. (1985) In Nuclear Magnetic Resonance of Liquid Crystals (Ed., Emsley, J.W.), Reidel, Dordrecht, The Netherlands, pp. 147–180.Google Scholar
  4. Emsley, J.W. and Lindon, J.C. (1975) NMR Spectroscopy using Liquid Crystal Solvents, Pergamon, New York, NY, U.S.A.Google Scholar
  5. Kung, H.C., Wang, K.Y., Goljer, I. and Bolton, P.H. (1995) J. Magn. Reson., B109, 323–325.Google Scholar
  6. Metz, G., Howard, K.P., Van Liemt, W.B.S., Prestegard, J.H., Lugtenburg, J. and Smith, S.O. (1995) J. Am. Chem. Soc., 117, 564–565.Google Scholar
  7. Ottiger, M., Tjandra, N. and Bax, A. (1997) J. Am. Chem. Soc., 119, in press.Google Scholar
  8. Prosser, R.S., Hunt, S.A., DiNatale, J.A. and Vold, R.R. (1996) J. Am. Chem. Soc., 118, 269–270.Google Scholar
  9. Salvatore, B.A., Ghose, R. and Prestegard, J.H. (1996) J. Am. Chem. Soc., 118, 4001–4008.Google Scholar
  10. Sanders, C.R. and Schwonek, J.P. (1992) Biochemistry, 31, 8898–8905.Google Scholar
  11. Sanders, C.R., Hare, B.J., Howard, K.P. and Prestegard, J.H. (1994) Prog. Nucl. Magn. Reson. Spectrosc., 26, 421–444.Google Scholar
  12. Schneider, D.M., Dellwo, M. and Wand, A.J. (1992) Biochemistry, 31, 3645–3652.Google Scholar
  13. Seelig, J., Borle, F. and Cross, T.A. (1985) Biochim. Biophys. Acta, {vn814} 195–198.Google Scholar
  14. Tjandra, N., Feller, S.E., Pastor, R.W. and Bax, A. (1995) J. Am. Chem. Soc., 117, 12562–12566.Google Scholar
  15. Tjandra, N., Grzesiek, S. and Bax, A. (1996) J. Am. Chem. Soc., 118, 6264–6272.Google Scholar
  16. Tjandra, N. and Bax, A. (1997) J. Magn. Reson., 124, 512–515.Google Scholar
  17. Tjandra, N., Omichinski, J.G., Gronenborn, A.M., Clore, G.M. and Bax, A. (1997) Nat. Struct. Biol., 4, 732–738.Google Scholar
  18. Tolman, J.R., Flanagan, J.M., Kennedy, M.A. and Prestegard, J.H. (1995) Proc. Natl. Acad. Sci. USA, 92, 9279–9283.Google Scholar
  19. Tolman, J.R. and Prestegard, J.H. (1996) J. Magn. Reson., B112, 245–252.Google Scholar
  20. Vijay-Kumar, S., Bugg, C.E. and Cook, W.J. (1987) J. Mol. Biol., {vn194}, 531–544.Google Scholar
  21. Vold, R.R. and Prosser, R.S. (1996) J. Magn. Reson., B113, 267–271.Google Scholar
  22. Wüthrich, K. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY, U.S.A.Google Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Ad Bax
    • 1
  • Nico Tjandra
    • 2
  1. 1.Laboratory of Chemical PhysicsNational Institute of Diabetes and Digestive and Kidney DiseasesBethesdaU.S.A
  2. 2.Laboratory of Biophysical ChemistryNational Heart, Lung, and Blood Institute, National Institutes of HealthBethesda

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