Clean SEA-HSQC: a method to map solvent exposed amides in large non-deuterated proteins with gradient-enhanced HSQC.
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.
Lin
Donghai
D
Department of Biochemistry, The Hong Kong University of Science and Technology, Kowloon, SAR, P R of China.
Sze
Kong Hung
KH
Cui
Yangfang
Y
Zhu
Guang
G
D000577:Amides / Q000737:chemistry*; D003903:Deuterium; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry*; D011522:Protons; D012997:Solvents / Q000737:chemistry*
D000577:Amides; D009587:Nitrogen Isotopes; D011506:Proteins; D011522:Protons; D012997:Solvents; D003903:Deuterium