CO_H(N)CACB experiments for assigning backbone resonances in 13C/15N-labeled proteins

Nathan Astrof, Clay Bracken, John Cavanagh, Arthur G. Palmer

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A triple resonance NMR experiment, denoted CO_H(N)CACB, correlates 1HN and 13CO spins with the 13Cα and 13Cβ spins of adjacent amino acids. The pulse sequence is an 'out-and-back' design that starts with 1HN magnetization and transfers coherence via the 15N spin simultaneously to the 13CO and 13Cα spins, followed by transfer to the 13Cβ spin. Two versions of the sequence are presented: one in which the 13CO spins are frequency labeled during an incremented t1 evolution period prior to transfer of magnetization from the 13Cα to the 13Cβ resonances, and one in which the 13CO spins are frequency labeled in a constant-time manner during the coherence transfer to and from the 13Cβ resonances. Because 13CO and 15N chemical shifts are largely uncorrelated, the technique will be especially useful when degeneracy in the 1HN-15N chemical shifts hinders resonance assignment. The CO_H(N)CACB experiment is demonstrated using uniformly 13C/15N-labeled ubiquitin.

Original languageEnglish
Pages (from-to)451-456
Number of pages6
JournalJournal of Biomolecular NMR
Volume11
Issue number4
DOIs
StatePublished - 1998
Externally publishedYes

Keywords

  • Constant time evolution
  • Heteronuclear NMR
  • Proteins
  • Sequential assignments
  • Triple resonance

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