Journal of Biomolecular NMR

, Volume 2, Issue 3, pp 257–274 | Cite as

Precise vicinal coupling constants3JHNα in proteins from nonlinear fits of J-modulated [15N,1H]-COSY experiments

  • Martin Billeter
  • Dario Neri
  • Gottfried Otting
  • Yan Qiu Qian
  • Kurt Wüthrich
Research Papers


Improved experimental schemes for the recently introduced J-modulated [15N,1H]-correlation experiment for measurements of the homonuclear amide proton-Cα proton vicinal coupling constants.3JHNα, in uniformly15N-labeled proteins are described, and a nonlinear fit procedure is presented for quantitative evaluation of3JHNα. The method was first tested with the N-terminal DNA-binding domain of the 434 repressor (M=7.3 kDa), where at 13 C precise values of3JHNα in the range 2.0–9.5 Hz were obtained for all residues with resolved15N-1H cross peaks. It was then applied to theAntennapedia homeodomain complexed to a synthetic 14-base pair DNA fragment (molecular weight of the complex ∼ 18 kDa). The3JHNα values measured were found to be in excellent agreement with those predicted from the secondary structure of this protein in the complex.


Vicinal spin-spin coupling constants J-modulated [15N,1H]-COSY Nonlinear fit of J-modulation Protein conformation NMR structure of proteins 

Abbreviations and symbols


nuclear Overhauser effect


two-dimensional correlated spectroscopy

3JHNα or J

homonuclear vicinal amide proton-Cα proton coupling constant

434 repressor(1–69)

N-terminal DNA-binding domain of the 434 repressor comprising 69 residues


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Copyright information

© ESCOM Science Publishers B.V 1992

Authors and Affiliations

  • Martin Billeter
    • 1
  • Dario Neri
    • 1
  • Gottfried Otting
    • 1
  • Yan Qiu Qian
    • 1
  • Kurt Wüthrich
    • 1
  1. 1.Institut für Molekularbiologie und BiophysikEidgenössische Technische Hochschule-HönggerbergZürichSwitzerland

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