Journal of Biomolecular NMR

, Volume 9, Issue 1, pp 94–100

(H)N(COCA)NH and HN(COCA)NH experiments for 1H-15N backbone assignments in 13C/15N-labeled proteins

  • Clay Bracken
  • Arthur G. Palmer III
  • John Cavanagh
Article

DOI: 10.1023/A:1018679819693

Cite this article as:
Bracken, C., III, A.G.P. & Cavanagh, J. J Biomol NMR (1997) 9: 94. doi:10.1023/A:1018679819693

Abstract

Triple resonance HN(COCA)NH pulse sequences for correlating 1H(i), 15N(i),1H(i-1), and 15N(i-1) spins that utilize overlapping coherence transfer periods provide increased sensitivityrelative to pulse sequences that utilize sequential coherence transfer periods. Although theoverlapping sequence elements reduce the overall duration of the pulse sequences, theprincipal benefit derives from a reduction in the number of 180° pulses. Two versions of thetechnique are presented: a 3D (H)N(COCA)NH experiment that correlates 15N(i),1H(i-1), and 15N(i-1) spins, and a 3D HN(COCA)NH experiment that correlates 1H(i), 15N(i),1H(i-1), and 15N(i-1) spins by simultaneously encoding the 1H(i) and 15N(i) chemical shiftsduring the t1 evolution period. The methods are demonstrated on a 13C/15N-enriched sampleof the protein ubiquitin and are easily adapted for application to 2H/13C/15N-enrichedproteins.

Triple resonance NMR spectroscopy Sequential assignment Proteins 

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Clay Bracken
    • 1
  • Arthur G. Palmer III
    • 1
  • John Cavanagh
    • 2
  1. 1.Department of Biochemistry and Molecular BiophysicsColumbia UniversityNew YorkU.S.A
  2. 2.NMR Structural Biology Facility, Wadsworth CenterNew York State Department of HealthAlbanyU.S.A

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