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Journal of Biomolecular NMR

, Volume 2, Issue 2, pp 203–210 | Cite as

A new 3D HN(CA)HA experiment for obtaining fingerprint HN-Hα cross peaks in15N- and13C-labeled proteins

  • Robert T. Clubb
  • V. Thanabal
  • Gerhard Wagner
Short Communications

Summary

A new 3D1H−15N−13C triple resonance experiment is presented that provides in-phase absorptive cross peaks between amide protons and α-protons of the same and the sequentially preceding residue. The experiment yields similar connectivities as those described previously by Montelione and Wagner (1990a) [J. Magn. Reson.,87, 183–188] and Kay et al. (1991) [J. Magn. Reson.,91, 84–92]. However, the pulse sequence was designed to minimize the time that transverse coherence of the13Cα nucleus is present, since this nucleus has the shortest transverse relaxation time of all the nuclei involved in these experiments. This is achieved by using a coherence transfer pathway from1HN to15N,13Cα,1Hα and back to the1HN. In the sequence described, transverse13Cα coherence is present only for a length of ca. I1J(Cα-Hα). This reduces loss of signal due to transverse relaxation. We tested the technique on uniformly15N- and13C-enriched T4 lysozyme.

Keywords

3D NMR J connectivity Resonance assignment Isotope labeling Proteins T4 lysozyme 

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Copyright information

© ESCOM Science Publishers B.V 1992

Authors and Affiliations

  • Robert T. Clubb
    • 1
    • 2
  • V. Thanabal
    • 1
  • Gerhard Wagner
    • 1
  1. 1.Department of Biological Chemistry and Molecular PharmacologyHarvard Medical SchoolBostonUSA
  2. 2.Department of Biological ChemistryUniversity of MichiganAnn ArborUSA

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