Advertisement

Journal of Biomolecular NMR

, Volume 31, Issue 4, pp 279–293 | Cite as

Residual backbone and side-chain 13C and 15N resonance assignments of the intrinsic transmembrane light-harvesting 2 protein complex by solid-state Magic Angle Spinning NMR spectroscopy

  • A. J. van Gammeren
  • F. B. Hulsbergen
  • J. G. Hollander
  • H. J. M. de GrootEmail author
Article

Abstract

This study reports the sequence specific chemical shifts assignments for 76 residues of the 94 residues containing monomeric unit of the photosynthetic light-harvesting 2 transmembrane protein complex from Rhodopseudomonas acidophila strain 10050, using Magic Angle Spinning (MAS) NMR in combination with extensive and selective biosynthetic isotope labeling methods. The sequence specific chemical shifts assignment is an essential step for structure determination by MAS NMR. Assignments have been performed on the basis of 2-dimensional proton-driven spin diffusion 13C–13C correlation experiments with mixing times of 20 and 500 ms and band selective 13C–15N correlation spectroscopy on a series of site-specific biosynthetically labeled samples. The decreased line width and the reduced number of correlation signals of the selectively labeled samples with respect to the uniformly labeled samples enable to resolve the narrowly distributed correlation signals of the backbone carbons and nitrogens involved in the long α-helical transmembrane segments. Inter-space correlations between nearby residues and between residues and the labeled BChl a cofactors, provided by the 13C–13C correlation experiments using a 500 ms spin diffusion period, are used to arrive at sequence specific chemical shift assignments for many residues in the protein complex. In this way it is demonstrated that MAS NMR methods combined with site-specific biosynthetic isotope labeling can be used for sequence specific assignment of the NMR response of transmembrane proteins.

Keywords

biosynthesis pattern isotope labeling transmembrane protein complex sequential assignment solid-state magic angle spinning NMR 

Abbreviations

LH2

light-harvesting 2 protein

PDSD

proton driven spin diffusion.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Alia Matysik, J., Soede-Huijbregts, C., Baldus, M., Raap, J., Lugtenburg, J., Gast, P., Gorkom, H.J., Hoff, A.J., Groot, H.J.M. 2001J. Am. Chem. Soc.12348034809CrossRefPubMedGoogle Scholar
  2. Baldus, M., Geurts, D.G., Hediger, S., Meier, B.H. 1996J. Magn. Reson. Ser. A118140144CrossRefGoogle Scholar
  3. Baldus, M., Petkova, A.T., Herzfeld, J., Griffin, R.G. 1998Mol. Phys.9511971207CrossRefGoogle Scholar
  4. Bennett, A.E., Rienstra, C.M., Griffiths, J.M., Zhen, W.G., Lansbury, P.T., Griffin, R.G. 1998J. Chem. Phys.10894639479Google Scholar
  5. Böckmann, A., Lange, A., Galinier, A., Luca, S., Giraud, N., Juy, M., Heise, H., Montserret, R., Penin, F., Baldus, M. 2003J. Biomol. NMR27323339PubMedGoogle Scholar
  6. Castellani, F., Rossum, A., Diehl, B., Rehbein, K., Oschkinat, H. 2003Biochemistry421147611483PubMedGoogle Scholar
  7. Castellani, F., Rossum, B., Diehl, A., Schubert, M., Rehbein, K., Oschkinat, H. 2002Nature42098102PubMedGoogle Scholar
  8. Detken, A., Hardy, E.H., Ernst, M., Kainosho, M., Kawakami, T., Aimoto, S., Meier, B.H. 2001J. Biomol. NMR20203221PubMedGoogle Scholar
  9. Egorova-Zachernyuk, T.A., Hollander, J., Fraser, N., Gast, P., Hoff, A.J., Cogdell, R.J., Groot, H.J.M., Baldus, M. 2001J. Biomol. NMR19243253PubMedGoogle Scholar
  10. Fujiwara, T., Todokoro, Y., Yanagishita, H., Tawarayama, M., Kohno, T., Wakamatsu, K., Akutsu, H. 2004J. Biomol. NMR28311325PubMedGoogle Scholar
  11. Hediger, S., Meier, B.H., Ernst, R.R. 1995Chem. Phys. Lett.240449456Google Scholar
  12. Hong, M. 1999Biophys. J.76A392A392Google Scholar
  13. Koepke, J., Hu, X.C., Muenke, C., Schulten, K., Michel, H. 1996Structure4581597PubMedGoogle Scholar
  14. Krogh, A., Larsson, B., Heijne, G., Sonnhammer, E.E.L. 2001J. Mol. Biol.305567580PubMedGoogle Scholar
  15. Kühlbrandt, W. 2001Nature411896899PubMedGoogle Scholar
  16. Marassi, F.M., Opella, S.J. 2003Protein Sci.12403411PubMedGoogle Scholar
  17. McDermott, G., Prince, S.M., Freer, A.A., Hawthornthwaite-Lawless, A.M., Papiz, M.Z., Cogdell, R.J., Isaacs, N.W. 1995Nature374517521Google Scholar
  18. Metz, G., Wu, X.L., Smith, S.O. 1994J. Magn. Reson. Ser. A110219227Google Scholar
  19. Opella, S.J., Marassi, F.M. 2004Chem. Rev.10435873606PubMedGoogle Scholar
  20. Opella, S.J., Nevzorov, A., Mesleh, M.F., Marassi, F.M. 2002Biochem. Cell Biol.80597604PubMedGoogle Scholar
  21. Palczewski, K., Kumasaka, T., Hori, T., Behnke, C.A., Motoshima, H., Fox, B.A., Le Trong, I., Teller, D.C., Okada, T., Stenkamp, R.E., Yamamoto, M., Miyano, M. 2000Science289739745CrossRefPubMedGoogle Scholar
  22. Papiz, M.Z., Prince, S.M., Howard, T., Cogdell, R.J., Isaacs, N.W. 2003J. Mol. Biol.32615231538PubMedGoogle Scholar
  23. Pauli, J., Baldus, M., Rossum, B., Groot, H.J.M., Oschkinat, H. 2001Chembiochem2272281PubMedGoogle Scholar
  24. Prince, S.M., Papiz, M.Z., Freer, A.A., McDermott, G., Hawthornthwaite-Lawless, A.M., Cogdell, R.J., Isaacs, N.W. 1997J. Mol. Biol.268412423PubMedGoogle Scholar
  25. Schulten, E.A.M., Matysik, J., Alia Kiihne, S., Raap, J., Lugtenburg, J., Gast, P., Hoff, A.J., Groot, H.J.M. 2002Biochemistry4187088717PubMedGoogle Scholar
  26. Simon, I., Fiser, A., Tusnady, G.E. 2001BBA-Protein Struct. M.1549123136Google Scholar
  27. Straus, S.K., Bremi, T., Ernst, R.R. 1998J. Biomol. NMR123950PubMedGoogle Scholar
  28. van Gammeren, A.J., Hulsbergen, F.B., Hollander, J.G., de␣Groot, H.J.M. 2004J. Biomol. NMR30267274PubMedGoogle Scholar
  29. van Gammeren, A.J., Buda, F., Hulsbergen, F.B., Kiihne, S., Hollander, J.G., Egorova-Zachernyuk, T.A., Fraser, N.J., Cogdell, R.J. and de Groot, H.J.M. (2004) J. Am. Chem. Soc. in pressGoogle Scholar
  30. Wallin, E., Heijne, G. 1998Protein Sci.710291038PubMedGoogle Scholar

Copyright information

© Springer 2005

Authors and Affiliations

  • A. J. van Gammeren
    • 1
  • F. B. Hulsbergen
    • 1
  • J. G. Hollander
    • 1
  • H. J. M. de Groot
    • 1
    Email author
  1. 1.Leiden Institute of ChemistryGorlaeus Laboratoria, Leiden UniversityThe Netherlands

Personalised recommendations