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Generation of serine/threonine check points in HN(C)N spectra

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Abstract

We describe here a simple modification of the HN(C)N experiment for the generation of serine/threonine check points in the three-dimensional experiment. The various ‘triplet of residue’ specific peak patterns in the spectra are documented for ease of analysis and sequential backbone resonance assignment. The performance of this experiment, referred to as HN(C)N-ST, is demonstrated using two proteins, one properly folded and the other completely denatured. It is noteworthy that, even in the denatured protein, where spectral dispersions are rather poor, about 90% of the sequential connectivities through the chain could be established from this single experiment. This would have great implications for structural genomics efforts.

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References

  1. Yee A, Gutmanas A and Arrowsmith C H 2006 Curr. Opin. Struct. Biol. 16 611

    Article  CAS  Google Scholar 

  2. Wuthrich K 1986 NMR of proteins and nucleic acids (New York: John Wiley & Sons)

    Google Scholar 

  3. Wagner G 1997 Natl. Struct. Biol. 4 Suppl. pp 841–844

  4. Bhavesh N S, Panchal S C and Hosur R V 2001 Biochemistry 40 14727

    Article  CAS  Google Scholar 

  5. Chatterjee A, Bhavesh N S, Panchal S C and Hosur R V 2002 Biochem. Biophys. Res. Commun. 293 427

    Article  CAS  Google Scholar 

  6. Chatterjee A, Kumar A and Hosur R V 2006 J. Magn. Reson. 181 21

    Article  CAS  Google Scholar 

  7. Chugh J, Kumar D and Hosur R V 2008 J. Biomol. NMR 40 145

    Article  CAS  Google Scholar 

  8. Bhavesh N S, Sinha R, Mohan P M and Hosur R V 2003 J. Biol. Chem. 278 19980

    Article  CAS  Google Scholar 

  9. Bhavesh N S, Juneja J, Udgaonkar J B and Hosur R V 2004 Protein Sci. 13 3085

    Article  CAS  Google Scholar 

  10. Chatterjee A, Mridula P, Mishra R K, Mittal R and Hosur R V 2005 J. Biol. Chem. 280 11369

    Article  CAS  Google Scholar 

  11. Chatterjee A, Krishna Mohan P M, Prabhu A, Ghosh-Roy A and Hosur R V 2007 Biochimie 89 117

    Article  CAS  Google Scholar 

  12. Chugh J, Chatterjee A, Kumar A, Mishra R K, Mittal R and Hosur R V 2006 FEBS J. 273 388

    Article  CAS  Google Scholar 

  13. Chugh J, Sharma S and Hosur R V 2007 Biochemistry 46 11819

    Article  CAS  Google Scholar 

  14. Chugh J, Sharma S, Kumar D and Hosur R V 2009 Biomolecular NMR Assignment

  15. Kumar A, Srivastava S, Kumar M R, Mittal R and Hosur R V 2006 J. Mol. Biol. 361 180

    Article  CAS  Google Scholar 

  16. Kumar A, Srivastava S, Mishra R K, Mittal R and Hosur R V 2006 Biophys. J. 90 2498

    Article  CAS  Google Scholar 

  17. Kumar D, Kumar A, Misra J R, Chugh J, Sharma S and Hosur R V 2008 1H, 15N, 13C resonance assignment of folded and 8 M urea denatured state of SUMO from Drosophila melanogaster pp. 13–15

  18. Barnwal R P, Jobby M K, Sharma Y and Chary K V 2006 J. Biomol. NMR 36 Suppl 1 32

    Article  Google Scholar 

  19. Chugh J, Sharma S, Kumar D and Hosur R V 2009 Biomolecular NMR Assignment 3 13–16

    Article  CAS  Google Scholar 

  20. Emsley L and Bodenhausen G 1990 Gaussian pulse cascades: New analytical functions for rectangular selective inversion and in-phase excitation in NMR pp. 469–476

  21. Panchal S C, Bhavesh N S and Hosur R V 2001 J. Biomol. NMR 20 135

    Article  CAS  Google Scholar 

  22. Piotto M, Saudek V and Sklenar V 1992 J. Biomol. NMR 2 661

    Article  CAS  Google Scholar 

  23. Marion D, Ikura M, Tschudin R and Bax A 1989 Application to the study of hydrogen exchange in proteins, pp. 393–399

  24. Grzesiek S and Bax A 1993 J. Am. Chem. Soc. 115 12593

    Article  CAS  Google Scholar 

  25. Bhavesh N S, Chatterjee A, Panchal S C and Hosur R V 2003 Biochem. Biophys. Res. Commun. 311 678

    Article  CAS  Google Scholar 

  26. Wirmer J and Schwalbe H 2002 J. Biomol. NMR 23 47

    Article  CAS  Google Scholar 

  27. BMRB DataBase 2009 http://www.bmrb.wisc.edu/ref_info/statful.htm#1

  28. Smith M A, Shaka A J and Hu H 2001 J. Magn. Reson. 151 269

    Article  CAS  Google Scholar 

  29. Silver M S, Joseph R I and Hoult D I 1985 Phys. Rev. A31 2753

    Google Scholar 

  30. Shaka A J, Keeler J and Freeman R 1983 Evaluation of a new broadband decoupling sequence: WALTZ-16 pp. 313–340

  31. Shaka A J, Keeler J, Frankiel T and Freeman R 1983 An improved sequence for broadband decoupling: WALTZ-16 pp. 335–338

  32. Shaka, A J, Barker P B and Freeman R 1985 Computer-optimized decoupling scheme for wideband applications and low-level operation pp. 547–552

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Authors and Affiliations

  1. Department of Chemical Sciences, Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai, 400 005, India

    Dinesh Kumar, Jeetender Chugh & Ramakrishna V. Hosur

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  1. Dinesh Kumar
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  2. Jeetender Chugh
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  3. Ramakrishna V. Hosur
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Correspondence to Ramakrishna V. Hosur.

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Both the authors have contributed equally

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Kumar, D., Chugh, J. & Hosur, R.V. Generation of serine/threonine check points in HN(C)N spectra. J Chem Sci 121, 955–964 (2009). https://doi.org/10.1007/s12039-009-0124-6

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  • DOI: https://doi.org/10.1007/s12039-009-0124-6

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