Journal of Biomolecular NMR

, Volume 5, Issue 3, pp 323–326

The use of heteronuclear cross-polarization for backbone assignment of 2H-, 15N- and 13C-labeled proteins: A pulse scheme for triple-resonance 4D correlation of sequential amide protons and 15N

  • Masahiro Shirakawa
  • Markus Wälchli
  • Masato Shimizu
  • Yoshimasa Kyogoku
Short Communications

DOI: 10.1007/BF00211761

Cite this article as:
Shirakawa, M., Wälchli, M., Shimizu, M. et al. J Biomol NMR (1995) 5: 323. doi:10.1007/BF00211761

Summary

A new four-dimensional pulse scheme is described for the main-chain assignment of proteins by means of the J connectivity of the amide proton and nitrogen resonances of adjacent residues. Since the new experiment, 4D CP-HN(COCA)NH, involves heteronuclear cross-polarization for magnetization transfer from 13C=O to 15N via 13Cα, a relatively strong WALTZ-16 decoupling rf field is applied to 13Cα during magnetization transfer. Consequently, 13Cα is effectively decoupled from its attached 2H in the case of deuterated proteins, in the absence of a decoupling rf field for 2H. This efficiently improves the sensitivity of the experiment through 13C line narrowing. The experiment was performed on a randomly 60% deuterated protein, and the sensitivity of the final 4D spectrum was found to be excellent.

Keywords

Heteronuclear NMRDeuterium labelingCross-polarizationTriple-resonance experimentPulse scheme

Copyright information

© ESCOM Science Publishers B.V. 1995

Authors and Affiliations

  • Masahiro Shirakawa
    • 1
  • Markus Wälchli
    • 2
  • Masato Shimizu
    • 1
  • Yoshimasa Kyogoku
    • 1
  1. 1.Institute for Protein ResearchOsaka UniversitySuita, Osaka 565Japan
  2. 2.Bruker JapanTsukuba, Ibaraki 305Japan