Sequential protein NMR assignments in the liquid state via sequential data acquisition

J Magn Reson. 2014 Feb:239:23-8. doi: 10.1016/j.jmr.2013.12.002. Epub 2013 Dec 14.

Abstract

Two different NMR pulse schemes involving sequential (1)H data acquisition are presented for achieving protein backbone sequential resonance assignments: (i) acquisition of 3D {HCCNH and HNCACONH} and (ii) collection of 3D {HNCOCANH and HNCACONH} chemical shift correlation spectra using uniformly (13)C,(15)N labelled proteins. The sequential acquisition of these spectra reduces the overall experimental time by a factor of ≈2 as compared to individual acquisitions. The suitability of this approach is experimentally demonstrated for the C-terminal winged helix (WH) domain of the minichromosome maintenance (MCM) complex of Sulfolobus solfataricus.

Keywords: Minichromosome maintenance (MCM); NMR; Protein resonance assignment; Sequential data acquisition; Winged helix domain (WH).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon Isotopes
  • Chromosomes, Bacterial / chemistry
  • Electromagnetic Fields
  • Indicators and Reagents
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Proteins / chemistry*
  • Sulfolobus solfataricus / chemistry

Substances

  • Carbon Isotopes
  • Indicators and Reagents
  • Nitrogen Isotopes
  • Proteins