Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in 15N-13C-ILV methyl labeled proteins

Dominique P. Frueh, Alison Leed, Haribabu Arthanari, Alexander Koglin, Christopher T. Walsh, Gerhard Wagner

Research output: Contribution to journalArticlepeer-review

Abstract

We present a time-shared 3D HSQC-NOESY experiment that enables one to simultaneously record 13C- and 15N-dispersed spectra in Ile, Leu and Val (ILV) methyl-labeled samples. This experiment is designed to delineate the two spectra which would otherwise overlap with one another when acquired together. These spectra display nOe correlations in the detected proton dimension, i.e. with maximum resolution. This is in contrast to NOESY-HSQC types of experiments that provide cross-peaks in the indirect dimension with low resolution due to limits in experimental time. The technique is particularly advantageous at high field where even longer experimental times would be required for comparable resolution in NOESY-HSQC experiments. The method is demonstrated at 900 MHz and at 750 MHz on 37 and 31 kDa proteins, respectively. The resolution and time saving provided in this experiment was crucial for solving the structures of these two proteins.

Original languageEnglish (US)
Pages (from-to)311-318
Number of pages8
JournalJournal of Biomolecular NMR
Volume45
Issue number3
DOIs
StatePublished - 2009
Externally publishedYes

Keywords

  • Distance constraints
  • High-resolution
  • NOESY
  • Non-ribosomal peptide synthetases
  • Nuclear magnetic resonance
  • Protein structure
  • Time-shared

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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