Semi-constant-time HMSQC (SCT-HMSQC-HA) for the measurement of (3)J(H(N))(H(alpha)) couplings in (15)N-labeled proteins

J Magn Reson. 2000 Apr;143(2):391-6. doi: 10.1006/jmre.1999.2010.

Abstract

A simple method for accurately measuring (3)J(H(N))(H(alpha)) coupling constants in (15)N-labeled proteins is described. This semi-constant-time HMSQC-HA experiment combines the rapidity and convenience of the recently introduced CT-HMQC-HA scheme (Postingl and Otting, J. Biomol. NMR 12, 319-324 (1998)) with the high resolution and robustness of the HSQC experiment. The proposed method is demonstrated for the 76-residue human ubiquitin and Saccharopolyspora erythraea calerythrin (176 residues). Our results imply that the SCT-HMSQC-HA experiment is suitable also for proteins with less favorable NMR properties due to its good resolution and sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / analysis
  • Bacterial Proteins / chemistry
  • Calcium-Binding Proteins / chemistry
  • Humans
  • Hydrogen
  • Hydrogen Bonding
  • Magnetic Resonance Spectroscopy / methods*
  • Nitrogen Isotopes
  • Proteins / chemistry*
  • Saccharopolyspora / chemistry
  • Sensitivity and Specificity
  • Spin Labels
  • Time Factors
  • Ubiquitins / chemistry

Substances

  • Amino Acids
  • Bacterial Proteins
  • Calcium-Binding Proteins
  • Nitrogen Isotopes
  • Proteins
  • Spin Labels
  • Ubiquitins
  • calerythrin protein, Saccharopolyspora erythraea
  • Hydrogen