Biomolecular NMR Assignments

, Volume 3, Issue 1, pp 29–31 | Cite as

1H, 15N and 13C assignments of domain 5 of Dictyostelium discoideum gelation factor (ABP-120) in its native and 8M urea-denatured states

  • Shang-Te Danny Hsu
  • Lisa D. Cabrita
  • John Christodoulou
  • Christopher M. Dobson
Article

Abstract

The gelation factor from Dictyostelium discoideum (ABP-120) is an actin binding protein consisting of six immunoglobulin (Ig) domains in the C-terminal rod domain. We have recently used the pair of domains 5 and 6 of ABP-120 as a model system for studying multi-domain nascent chain folding on the ribosome. Here we present the NMR assignments of domain 5 in its native and 8M urea-denatured states.

Keywords

Protein folding Nascent chain Urea Ribosome ABP-120 

Notes

Acknowledgements

STDH is a recipient of a Netherlands Ramsay and a Human Frontier Science Program Long-term Fellowship (LT0798/2005). LDC is a NH&MRC C.J. Martin Fellow. JC is a recipient of an HFSP Young Investigator’s Award (RGY67/2007). JC and CMD acknowledge funding from the Wellcome and Leverhulme Trusts. We thank Prof. Paola Fucini for providing the DNA plasmid of domains 5 and 6 of ABP-120 from which the current construct was derived, and the staff of the Biomolecular NMR Facility, Department of Chemistry, University of Cambridge.

References

  1. Delaglio F, Grzesiek S, Vuister GW, Zhu G, Pfeifer J, Bax A (1995) NMRPipe: a multidimensional spectral processing system based on UNIX pipes. J Biomol NMR 6:277–293. doi: 10.1007/BF00197809 CrossRefGoogle Scholar
  2. Gilbert RJC, Fucini P, Connell S, Fuller SD, Nierhaus KH, Robinson CV, Dobson CM, Stuart DI (2004) Three-dimensional structures of translating ribosomes by cryo-EM. Mol Cell 14:57–66. doi: 10.1016/S1097-2765(04)00163-7 CrossRefGoogle Scholar
  3. Goddard TD, Kneller DG Sparky 3. University of California: San Francisco. http://www.cgl.ucsf.edu/home/sparky/
  4. Hsu S-TD, Fucini P, Cabrita LD, Launay H, Dobson CM, Christodoulou J (2007) Structure and dynamics of a ribosome-bound nascent chain by NMR spectroscopy. Proc Natl Acad Sci USA 104:16516–16521. doi: 10.1073/pnas.0704664104 CrossRefADSGoogle Scholar
  5. Jung YS, Zweckstetter M (2004) Mars—robust automatic backbone assignment of proteins. J Biomol NMR 30:11–23. doi: 10.1023/B:JNMR.0000042954.99056.ad CrossRefGoogle Scholar
  6. McCoy AJ, Fucini P, Noegel AA, Stewart M (1999) Structural basis for dimerization of the Dictyostelium gelation factor (ABP120) rod. Nat Struct Biol 6:836–841. doi: 10.1038/12296 CrossRefGoogle Scholar
  7. Popowicz GM, Muller R, Noegel AA, Schleicher M, Huber R, Holak TA (2004) Molecular structure of the rod domain of Dictyostelium filamin. J Mol Biol 342:1637–1646. doi: 10.1016/j.jmb.2004.08.017 CrossRefGoogle Scholar
  8. Popowicz GM, Schleicher M, Noegel AA, Holak TA (2006) Filamins: promiscuous organizers of the cytoskeleton. Trends Biochem Sci 31:411–419. doi: 10.1016/j.tibs.2006.05.006 CrossRefGoogle Scholar
  9. Schwaiger I, Kardinal A, Schleicher M, Noegel AA, Rief M (2004) A mechanical unfolding intermediate in an actin-crosslinking protein. Nat Struct Mol Biol 11:81–85. doi: 10.1038/nsmb705 CrossRefGoogle Scholar
  10. Schwarzinger S, Kroon GJ, Foss TR, Chung J, Wright PE, Dyson HJ (2001) Sequence-dependent correction of random coil NMR chemical shifts. J Am Chem Soc 123:2970–2978. doi: 10.1021/ja003760i CrossRefGoogle Scholar
  11. van der Flier A, Sonnenberg A (2001) Structural and functional aspects of filamins. Biochim Biophys Acta 1538:99–117. doi: 10.1016/S0167-4889(01)00072-6 CrossRefGoogle Scholar
  12. Wishart DS, Bigam CG, Yao J, Abildgaard F, Dyson HJ, Oldfield E, Markley JL, Sykes BD (1995) 1H, 13C and 15N chemical shift referencing in biomolecular NMR. J Biomol NMR 6:135–140. doi: 10.1007/BF00211777 CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Shang-Te Danny Hsu
    • 1
  • Lisa D. Cabrita
    • 1
    • 2
  • John Christodoulou
    • 1
    • 2
  • Christopher M. Dobson
    • 1
  1. 1.Department of ChemistryUniversity of CambridgeCambridgeUnited Kingdom
  2. 2.Institute of Structural and Molecular Biology, Research Department of Structural and Molecular Biology, UCL and School of Crystallography, Birkbeck CollegeUniversity of LondonLondonUnited Kingdom

Personalised recommendations