Journal of Biomolecular NMR

, Volume 36, Issue 1, pp 13–26

Towards unambiguous assignment of methyl-containing residues by double and triple sensitivity-enhanced HCCmHm-TOCSY experiments

Authors

  • Peter Würtz
    • Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR LaboratoryUniversity of Helsinki
  • Maarit Hellman
    • Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR LaboratoryUniversity of Helsinki
  • Helena Tossavainen
    • Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR LaboratoryUniversity of Helsinki
    • Program in Structural Biology and Biophysics, Institute of Biotechnology/NMR LaboratoryUniversity of Helsinki
Article

DOI: 10.1007/s10858-006-9056-3

Cite this article as:
Würtz, P., Hellman, M., Tossavainen, H. et al. J Biomol NMR (2006) 36: 13. doi:10.1007/s10858-006-9056-3

Abstract

Chemical shift assignment of methyl-containing residues is essential in protein NMR spectroscopy, as these residues are abundant in protein interiors and provide the vast majority of long-range NOE connectivities for structure determination. These residues also constitute an integral part of hydrophobic cavities, the surroundings for many enzymatic reactions. Here we present a powerful strategy for the assignment of methyl-containing residues in a uniformly 13C/15N double labeled protein sample. The approach is based on novel four-dimensional HCCmHm-TOCSY experiments, two of them utilizing gradient selection and sensitivity enhancement in all three indirectly detected dimensions. Regardless of the number of dimensions, the proposed experiments can be executed using only one transient per FID, providing outstanding resolution and sensitivity. A complete assignment of the 51 methyl-containing residues in the 16 kDa Mus musculus coactosin was accomplished using a four-dimensional HCCmHm-TOCSY spectrum recorded in 16 hours.

Keywords

assignmentcoactosincoherence transferHCCH-TOCSYmethyl groupsNMR spectroscopyproteins

Copyright information

© Springer Science+Business Media B.V. 2006