Protein backbone motions viewed by intraresidue and sequential HN–Hα residual dipolar couplings

  • Beat Vögeli
  • Lishan Yao
  • Ad BaxEmail author


Triple resonance E.COSY-based techniques were used to measure intra-residue and sequential HN–Hα residual dipolar couplings (RDCs) for the third IgG-binding domain of protein G (GB3), aligned in Pf1 medium. Measurements closely correlate with values predicted on the basis of an NMR structure, previously determined on the basis of a large number of one-bond backbone RDCs measured in five alignment media. However, in particular the sequential HN–Hα RDCs are smaller than predicted for a static structure, suggesting a degree of motion for these internuclear vectors that exceeds that of the backbone amide N–H vectors. Of all experimentally determined GB3 structures available, the best correlation between experimental 1H–1H couplings is observed for a GB3 ensemble, previously derived to generate a realistic picture of the conformational space sampled by GB3 (Clore and Schwieters, J Mol Biol 355:879–886, 2006). However, for both NMR and X-ray-derived structures the 1H–1H couplings are found to be systematically smaller than expected on the basis of alignment tensors derived from 15N–1H amide RDCs, assuming librationally corrected N–H bond lengths of 1.041 Å.


1H–1H dipolar coupling Backbone dynamics GB3 Residual dipolar coupling RDC 



We thank Drs. Lisa Parsons for help in sample preparation, and Dennis A. Torchia and G. Marius Clore for helpful discussions. Financial support was obtained from the Swiss National Science Foundation (to B.V.). This work was supported by the Intramural Research Program of the NIDDK, NIH, and by the Intramural AIDS-Targeted Antiviral Program of the Office of the Director, NIH.

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Copyright information

© US Government 2008

Authors and Affiliations

  1. 1.Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney DiseasesNational Institutes of HealthBethesdaUSA

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