Clean SEA-HSQC: a method to map solvent exposed amides in large non-deuterated proteins with gradient-enhanced HSQC

J Biomol NMR. 2002 Aug;23(4):317-22. doi: 10.1023/a:1020225206644.

Abstract

The recent introduction of the SEA-TROSY experiment (Pellecchia et al. (2001) J. Am. Chem. Soc., 123, 4633-4634) can alleviate the problem of resonance overlap in 15N/2H labeled proteins. This method selectively observes solvent exposed amide protons with a SEA element. However, SEA-TROSY spectra may be contaminated with exchange-relayed NOE contributions from fast exchanging hydroxyl or amine protons and longitudinal relaxation contributions. Furthermore, for non-deuterated proteins or protein-ligand complexes, SEA-TROSY spectra may contain NOE contributions from aliphatic protons. In this communication, a modified version of the SEA element, a Clean SEA element, is introduced to eliminate these artifacts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides / chemistry*
  • Deuterium
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Peptide Mapping
  • Proteins / chemistry*
  • Protons
  • Solvents / chemistry*

Substances

  • Amides
  • Nitrogen Isotopes
  • Proteins
  • Protons
  • Solvents
  • Deuterium