CACA-TOCSY with alternate 13C-12C labeling: A 13Cα direct detection experiment for mainchain resonance assignment, dihedral angle information, and amino acid type identification

Koh Takeuchi, Dominique P. Frueh, Zhen Yu J. Sun, Sebastian Hiller, Gerhard Wagner

Research output: Contribution to journalArticlepeer-review

Abstract

We present a 13C direct detection CACA-TOCSY experiment for samples with alternate 13C-12C labeling. It provides inter-residue correlations between 13Cα resonances of residue i and adjacent Cαs at positions i - 1 and i + 1. Furthermore, longer mixing times yield correlations to Cα nuclei separated by more than one residue. The experiment also provides C α-to-sidechain correlations, some amino acid type identifications and estimates for ψ dihedral angles. The power of the experiment derives from the alternate 13C-12C labeling with [1,3-13C] glycerol or [2-13C] glycerol, which allows utilizing the small scalar 3JCC couplings that are masked by strong 1JCC couplings in uniformly 13C labeled samples.

Original languageEnglish (US)
Pages (from-to)55-63
Number of pages9
JournalJournal of Biomolecular NMR
Volume47
Issue number1
DOIs
StatePublished - May 2010

Keywords

  • Alternate C labeling
  • C direct detection
  • Dihedral angle
  • Nuclear magnetic resonance (NMR)
  • Resonance assignment
  • TOCSY

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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